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Download Volume 11 - American Academy of Forensic Sciences
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AMERICAN ACADEMY OF FORENSIC SCIENCES 410 North 21st Street PO Box 669 Colorado Springs, CO 80901-0669 Phone: (719) 636-1100 Fax: (719) 636-5245 E-mail: [email protected] Website: www.aafs.org PROCEEDINGS of the American Academy of Forensic Sciences The Proceedings of the American Academy of Forensic Sciences is an official publication of the American Academy of Forensic Sciences (AAFS). It is devoted to the publication of the abstracts of technical oral papers and posters presented at the AAFS annual meeting. These include various branches of the forensic sciences such as pathology, toxicology, physical anthropology, psychiatry, immunology, odontology, jurisprudence, criminalistics, questioned documents, and engineering. Similar submissions dealing with forensic oriented aspects of the social sciences are also included. Please note that some of the abstracts included in the Proceedings deal with topics, results, and/or conclusions which are controversial. The publication of abstracts does not imply that the AAFS, its sections, or the individual section program chairs/committee has verified or agrees with the studies, results, and/or conclusions of each abstract. During the process of planning a scientific program, it is impossible to “peer-review” each abstract and presentation to the degree that is accomplished during manuscript review. Abstracts and presentations are accepted, in part, so that they can be critiqued and reviewed by other scientists, and so that a forum is available to discuss controversial issues. The views expressed in this publication are not those of the AAFS. The data and opinions appearing in the published material were prepared by and are the responsibility of the contributor(s), not of AAFS or its respective employees, employers, officers, and agents. The AAFS does not supply copies of meeting papers. It is suggested that you write directly to individual authors to obtain copies of specific papers. Presentation of some abstracts may have been scheduled or canceled after the publication of this document. English is the official language of the AAFS and its meetings; neither oral nor written translations will be provided. Copyright 2005 by the AAFS. Unless stated otherwise, noncommercial photocopying of editorial material published in this periodical is permitted by AAFS. Permission to reprint, publish, or otherwise reproduce such material in any form other than photocopying must be obtained from AAFS. Printed in the United States of America by Publication Printers, Corp., Denver, CO. PROCEEDINGS of the American Academy of Forensic Sciences Contents Financial Disclosure Index ...........................................................1 Special Sessions ..........................................................................10 Breakfast Seminars .....................................................................12 Luncheon Seminars.....................................................................17 Workshops & Workshorts ...........................................................18 Scientific Sessions Criminalistics.........................................................................32 Engineering Sciences...........................................................123 General.................................................................................159 Jurisprudence .......................................................................200 Odontology ..........................................................................210 Pathology/Biology ...............................................................227 Physical Anthropology ........................................................282 Psychiatry & Behavioral Science ........................................338 Questioned Documents........................................................347 Toxicology ...........................................................................357 Last Word.............................................................................377 Presenting Author Index ...........................................................382 Key Word Index ........................................................................387 February 2005 Volume XI Financial Disclosure As a sponsor of continuing education, the American Academy of Forensic Sciences must insure balance, independence, objectivity, and scientific rigor in all its educational activities. All faculty participating in a sponsoring activity are expected to disclose to the activity audience any significant financial interest or other relationship: (1) with the manufacturer(s) of any commercial product(s) and/or provider(s) of commercial services discussed in an educational presentation, and (2) with any commercial supporters of the activity. (Significant financial interest or other relationship can include such things as grants or research support, employee, consultant, major stockholder, member of speaker’s bureaus, etc.) The intent of this disclosure is not to prevent a speaker with a significant financial or other relationship from making a presentation, but rather to provide listeners with information on which they can make their own judgments. It remains for the audience to determine whether the speaker’s interest or relationships may influence the presentation with regard to the exposition or conclusion. The executed Financial Disclosure Forms are on file in the AAFS Office. As an accredited provider of Continuing Medical Education, the American Academy of Forensic Sciences is required to ask speakers to disclose any real or apparent conflict of interest they may have as related to the contents of their presentation(s). The existence of commercial or financial interests of authors related to the subject matter of their presentation(s) should not be construed as implying bias or decreasing the value of their presentation(s), however; disclosure should help participants form their own judgments. Authors not listed in this index indicated no affiliations with or financial interests in any commercial organizations. Those authors who disclosed affiliations or financial interests with the commercial organizations involved with products, to which they may refer, are listed below. A Ivo Alberink, PhD - D15 FearID Project (European Union Project Number G6RD-CT-200100618) (Grant/Research Support) Boyd Allen, MASc - C2 Accident Reconstruction Computer Software José R. Almirall, PhD - W10 NITECRIME (European Union Funded Network) (Grant/Research Support) Agilent New Wave (Demonstration Equipment) Joseph Almog, PhD - B161 Ferrotrace Mistral TSWG (Grant/Research Support) Michelle Alvarez, BS - B171 Technical Support Working Group (TSWG) on Counterterrorism (Grant/Research Support) Donald J. Anderson, BS - W7 Stark rxp (Other Financial/Material Support) Robert N. Anderson, PhD, PE - W7 Stark rxp (Other Financial/Material Support) B Erin K. Bai, BA - K11 California Department of Justice, BFS, Toxicology Laboratory (Other Financial/Material Support) Cerex Polycrom Clin II Solid Phase Extraction Column SPEware James A. Bailey, PhD - D75 Kodak Polychotrast RC Type F Photographic Paper Lora Bailey-Van Houten, BS - B1 California State University (Grant/Research Support) Joan E. Baker, PhD - H36 Joint POW/MIA Accounting Command (Employee) Robert E. Barsley, DDS, JD - SS2 Florida International University (Other Financial/Material Support) Joel Bartholf, BS - W16 3SI Security Systems (Employee) 3SI Security Systems FlexPac™ 3SI Security Systems Octapus™ 3SI Security Systems Scorpion®/Currency Guard United States Currency Protection Corporation Rachel A. Bartholomew, PhD - B2 Agilent 2100 Bioanalyzer Qiagen RNAEasy Micro Kits Qiagen Sensiscript Reverse Transcriptor Edward G. Bartick, PhD - B152 Federal Bureau of Investigation (Grant/Research Support) Permount Norland Optreal Adhesive (provided by University of South Carolina/Federal Bureau of Investigation) Martin Bauer, MD - B70 Applied Biosystems, Inc. Taqman® Gene Expression Assay University of Wuerzburg (Grant/Research Support) Adam C. Becnel, BS - SS2 Florida International University (Other Financial/Material Support) Philip S.L. Beh, MBBS, DMJ - G92 HKU CRCG (Grant/Research Support) RGC (Grant/Research Support) Peter J. Belcastro, Jr., MFS - J19 Federal Bureau of Investigation Laboratory (Speaker’s Bureau) Suzanne Bell, PhD - W3 Prentice Hall/Pearson Education (Other Financial/Material Support) Jason Berger, MS - B68 Applied Biosystems, Inc. ABI Prism® 3100 Genetic Analyzer Promega PowerPlex®16 Joan M. Bienvenue, MS Federal Bureau of Investigation (Grant/Research Support) – B78, B100 National Institute of Health (Grant/Research Support) – B78 Applied Biosystems, Inc. ABI 310 Genetic Analyzer – B100 Applied Biosystems, Inc. COfiler® - B100 Applied Biosystems, Inc. Profiler® Kits – B100 Applied Biosystems, Inc. POP-4 Polymer – B100 Sherie A. Blackwell, BSc - F29 University of Melbourne School of Dental Science (Employee) Laser Scanning (provided by Scanning and Inspection Pty., Ltd.) Herbert L. Blitzer, AB, MBA - D32 National Institute of Justice (Grant/Research Support) Cherisse Boland - B80 Applied Biosystems, Inc. (Employee) Applera Corporation AB Design Applera Corporation iscience Applera Corporation iscience Design Applera Corporation FAM® Applera Corporation NED® Applera Corporation PET® Applera Corporation Quantifiler™ Applera Corporation YFiler™ Applied Biosystems, Inc. ABI Prism® Applied Biosystems, Inc. AmpFlSTR® Identifiler™ Applied Biosystems, Inc. GeneScan® Applied Biosystems, Inc. GeneMapper® Applied Biosystems, Inc. Genotyper® Applied Biosystems, Inc. LIZ® Applied Biosystems, Inc. VIC® 1 Susan A. Bollinger, DDS - F7 Adobe® Photoshop® Richard S. Brown, MS - C45 MVA Scientific Consultants (Employee) Light and Electron Microscopy (provided by MVA Scientific Consultants) Sherry Brown, MS - W3 Prentice Hall/Pearson Education (Other Financial/Material Support) Aaron L. Brudenelle, MS - W3 Prentice Hall/Pearson Education (Other Financial/Material Support) Cynthia Brzozowski, DMD - F22 Adobe® Photoshop® Microsoft® Access Microsoft® Visual Basic WinBite Rebecca A. Bullard, BA, MS - G53 University of California, Davis (Grant/Research Support) JoAnn Buscaglia, PhD - W10 NITECRIME (European Union Funded Network) (Grant/Research Support) Agilent New Wave (Demonstration Equipment) C Pamela A. Cadiente - D46 Acuvue® Contact Lenses Baush & Laumb Contact Lenses Focus Dailies® Contact Lenses Chelex® (provided by Applied Biosystems, Inc.) USDE Ronald E. McNair Scholars Summer Research Institute Grant P217A030070 (Grant/Research Support) Lisa M. Calandro, MPH - B143 Applied Biosystems, Inc. (Employee) Applera Corporation AB Design Applera Corporation FAM® Applera Corporation iscience Applera Corporation iscience Design Applera Corporation NED® Applera Corporation PET® Applera Corporation Quantifiler™ Applera Corporation Yfiler™ Applied Biosystems, Inc. Genotyper® Applied Biosystems, Inc. GeneScan® Applied Biosystems, Inc. GeneMapper® Applied Biosystems, Inc. ABI Prism® Applied Biosystems, Inc. AmpFlSTR® Applied Biosystems, Inc. Identifiler™ Applied Biosystems, Inc. LIZ® Applied Biosystems, Inc. VIC® Carlo P. Campobasso, MD, PhD - G45 Doctoral Program in “Forensic Pathology and Criminalistics Techniques” (Grant/Research Support) Theresa A. Caragine, PhD - B71 Office of the Chief Medical Examiner of the City of New York (Other Financial/Material Support) Titan Inc. (Other Financial/Material Support) Beckman-Coulter Biomek 2000 Robot Promega DNA IQ™ Strip Testing (provided by Raven Laboratories) Eric J. Carita, MS - B107 NIJ (Grant/Research Support) Applied Biosystems, Inc. COfiler® STR Kits Applied Biosystems, Inc. Genescan® Applied Biosystems, Inc. Genotyper® Applied Biosystems, Inc. Profiler Plus® 2 John M. Carson, DDS - F13 Lumen Software Forensic IQ Joe S. Cecil, PhD, JD - W7 Stark rxp (Other Financial/Material Support) Mario J. Cellarosi, BA, MS - C58 National Institute of Standards and Technology (Employee) Salih Cengiz, PhD - B86 Research fund of Istanbul University (Grant/Research Support) Chien-Wei Chang, PhD - B16 Applied Biosystems, Inc. (Employee) Applera Corporation Quantifiler™ Real-Time PCR Kits Applied Biosystems, Inc. AmpFlSTR® Yfiler™ Kit Konstantinos H. Cherpelis, DDS - F18 WinID (provided by the OCME City of New York) Adobe® Photoshop® (provided by the OCME City of New York) Helen Cho, PhD - W2 Leica Microsystems Microscopes (Demonstration Equipment) Steven C. Clark, PhD - D40 National Association of Medical Examiners (Consultant) Brandi L. Clelland - D70 Federal Bureau of Investigation (Grant/Research Support) John T. Cody, PhD - K3 Dexedrine United Chemical Technologies, Inc XRDAH203 High Flow 200-mg Columns United Chemical Technologies, Inc XtracktT® Kenneth Cohrn, DDS - F27 Adobe® Photoshop® Edward J. Cone, PhD - K29 OraSure Technologies (Consultant) Oral Fluid Testing Device (provided by OraSure Technologies) Laura M. Conner, BS - D21 Bacharach TLV Sniffer Portable Arson Sampler tpi Pocket Combustable Gas Leak Detector Gail A. Cooper, PhD - K15 Cozart Biosciences, Ltd. (Employee) Cozart EIA Cannabinoids Microplate Kit Gilbert E. Corrigan, MS, MD, PhD - G64 Microsoft®: Word Microsoft® Visio Sulekha R. Coticone, PhD - B96 California State University Fresno Seed Grant (Grant/Research Support) Applied Biosystems, Inc. AmpFlSTR® Profiler Plus®ID™ Applied Biosystems, Inc. GeneMapper®ID Katie L. Coy, BS - B103 Applied Biosystems, Inc. AmpFlSTR® Profiler Plus® Natasha E. Coyle, MSc, PhD - D51 Applied Biosystems, Inc. (Employee) Applera Corporation PET® Applied Biosystems, Inc. ABI Prism® Genetic Analyzer Applied Biosystems, Inc. AmpFlSTR® Applied Biosystems, Inc. Identifiler® Applied Biosystems, Inc. PCR Amplification Kit Applied Biosystems, Inc. VIC® Christine L. Craig, MS - B32 ESDA2 Foster and Freeman Electrostatic Dust Print Lifter Sirchie David R. Craig, BS - C40 The Gas Technology Institute (Grant/Research Support) Rhonda Craig, MS - B102 Clorox® Bleach Fluorescein (provided by Gabor Patonay/Georgia State University) JOY® Dishwashing Detergent Sharon R. Crowley, RN, MN, FCNS - G83 CooperSurgical/Leisegang, Inc. (Other Financial/Material Support) Leisegang Colposcope CooperSurgical/Leisegang, Inc. D Jon C. Dailey, DDS - F10 ToothPrints® (provided by Kerr Dental) Marie Danforth, PhD, MA, AB - H63 University of Southern Mississippi (Grant/Research Support) Jon M. Davoren, MS - B134 International Commission on Missing Persons - ICMP (Employee) Applera Corporation Quantifiler™ Promega PowerPlex®16 System SeFiler Kits (provided by ICMP) Nunzio Di Nunno, MD, PhD - G29 Bari University (Grant/Research Support) Promega SV Total RNA Isolation Giancarlo Di Vella, MD, PhD - G76 .25 Caliber Baby Browning BMW Taylor M. Dickerson, MS - B67 Applied Biosystems, Inc. ABI Prism® 3100 Genetic Analyzer Bio-Rad 5% Chelex® Promega DNA IQ™ Promega PowerPlex® 16 Qiagen QIAamp™ James J. DiSarno, MS - SS2 Florida International University (Other Financial/Material Support) J.C. Upshaw Downs, MD - G80 .17 HMR round CCI Ammunition – G79 Century International Arms Tokarev CZ52 7.62 Pistol - G80 Jason P. Doyle, MFS - D62 NCIS Forensic Consultant Division (Employee) Spex Forensics Mini-Crimescope 400 Matthew J. Ducote, PhD - B11 Federal Bureau of Investigation (Grant/Research Support) Bio101® Soil DNA Extraction Kits BioVentures, Inc. Soil DNA Extraction Kits Epicentre® Soil DNA Extraction Kits MoBio Laboratories Soil DNA Extraction Kits Qiagen Soil DNA Extraction Kits Diana A. Dupuis, BA - H3 Forensic Sciences Foundation (Grant/Research Support) E Gavin R. Edmondstone, BSc - B57 Centre of Forensic Sciences (Employee) Jim Ehleringer, PhD - W10 NITECRIME (European Union Funded Network) (Grant/Research Support) Agilent (Demonstration Equipment) New Wave (Demonstration Equipment) Lim Eng Seng Simon, BS - B144 Applied Biosystems, Inc. Identifiler™ Applied Biosystems, Inc. Profiler Plus® Reliagene Technologies, Inc. Y-Plex™12 Heather L. Erek, BS - B106 Orchid Cellmark (Employee) Max B. Erwine, BS - W15 SPE Products & Techniques (Unlabeled Pharmaceutical/Investigational/ Medical Device) Varian, Inc. (Employee) Varian, Inc. SPE Phases * Presenting Author Sheila M. Estacio, BA NYC OCME (Employee) – B146 Florida International University (Other Financial/Material Support) – SS2 Applied Biosystems, Inc. ABI Prism® 3100 Genetic Analyzer – B146 Applied Biosystems, Inc. Quantiblot – B146 Bio RAD Micro Bio-Spin Chromatography Columns – B146 Promega DNA IQ – B146 Promega PowerPlex®16 – B146 F Lisa Faber, MFS - B63 NIJ No-Suspect Case DNA Backlog Reduction Program (Grant/Research Support) David R. Fisher, BS - B128 Roche Linear Array Mitochondrial DNA HVI/HVII RegionSequence Typing Kit Erica L. Fleishaker - B147 NIJ 2002-IJ-CX-K007 (Grant/Research Support) Corbett Research Rotor-Gene RG3000 Millipore Microcon® YM-30 Carolyn J. Fowler, PhD, MPH - D61 Microsoft® SQL Software (provided by Johns Hopkins Bloomberg School of Public Health) Bethany G. Fox - B14 P217A030070 (Grant/Research Support) Donna Freid, MA - H20 FORDISC Computer Software (provided by University of Tennessee, Knoxville) Amanda K. Frohwein, BS - SS2 Florida International University (Other Financial/Material Support) Michael E. Frontz, MSFS - K13 Bexar County Medical Examiner’s Office (Employee) Pennzoil-Quaker State Snap Brake Fluid G Robert E. Gaensslen, PhD - W3 Prentice Hall/Pearson Education (Other Financial/Material Support) Zeno J. Geradts, PhD – C19, D17 Ministry of Justice (Employee) Eugene Giles, PhD - H21 FORDISC 2.0 (provided by Forensic Anthropology Center) Ingrid A. Gill, JD Ancestry (provided by DNA DNAPrint Genomics, Inc.) - BS1 Applied Biosystems, Inc. - E4 Bode Technologies - E4 DNAPrint Genomics - E4 Forensic Science Associates - E4 Mitotyping Technologies - E4 Orchid Cellmark - E4 Promega - E4 ReliaGene, Inc. - E4 Technical Associates - E4 Justin M. Godby, BS - G54 MUFSC, WVDHHR/DSDC/OLS (Grant/Research Support) NASBA (provided by WVDHHR/DSC/OLS) Kenneth W. Goddard, MS - SS2 Florida International University (Other Financial/Material Support) Christopher M. Gojcz, BS - SS2 Florida International University (Other Financial/Material Support) Julie E. Golden, MS - B97 Applied Biosystems, Inc., Inc. ABI Prism® 310 Genetic Analyzer Applied Biosystems, Inc., Inc. AmpFlSTR® Applied Biosystems, Inc., Inc. Cofier/Profiler Plus® 3 Eleanor A.M. Graham, MSc - B72 National Society for the Prevention of Cruelty to Children, UK (Grant/Research Support) Applied Biosystems, Inc., Inc. ABI Prism® 377 DNA Sequencer Applied Biosystems, Inc., Inc. AmpFlSTR® SGM+® PCR Amplification Kit Applied Biosystems, Inc. GeneScan® Applied Biosystems, Inc. Genotyper® Bio-Rad Laboratories Chelex® 100 Resin Molecular Probes Oligreen® ssDNA Martin Grassberger, MD, PhD - G3 Agrobiogen TypiFix System ™Tags Phillip H. Grimm, BS - W16 United States Currency Protection Corporation (Employee) 3SI Security Systems FlexPac™ 3SI Security Systems Octapus™ 3SI Security Systems Scorpion®/Currency Guard H Ashley M. Hall, MS - B170 National Institute of Justice, University of Central Florida (Grant/Research Support) Joy L. Halverson, DVM - D58 DNA Typing Services QuestGen Forensics David B. Harte, DMD - D26 Toothprints® (provided by Kerr Corporation) Lotte Henke, PhD - B164 Promega PowerPlex®16 Katherine Henkelman - B108 Cedar Crest College (Grant/Research Support) Promega SE33 Kit Lori K. Hennessy, PhD - B142 Applied Biosystems, Inc. (Employee) Applera Corporation AB Design Applera Corporation FAM® Applera Corporation iscience Applera iscience Design Applera Corporation NED® Applera Corporation PET® Applera Corporation Quantifiler ™ Applera Corporation Yfiler™ Applied Biosystems, Inc. ABI Prism® Applied Biosystems, Inc. AmpFLSTR® Applied Biosystems, Inc. Identifiler® Applied Biosystems, Inc. Genotyper® Applied Biosystems, Inc. GeneScan® Applied Biosystems, Inc. GeneMapper® Applied Biosystems, Inc. LIZ™ Applied Biosystems, Inc. PCR Amplification Kit Applied Biosystems, Inc. VIC® Nicholas Hermann, PhD - W20 FORDISC (provided by the University of Tennessee) SAS (provided by the University of Tennessee) SYSTAT (provided by the University of Tennessee) Jeffrey A. Hickey, MS - B138 Applied Biosystems, Inc. Profiler Plus® Applied Biosystems, Inc. Taqman® Promega PowerPlex® Y Ashley J. Hinkle, BS - B95 Marshall University Forensic Science Center (Grant/Research Support) Applied Biosystems, Inc. ABI 7000 Sequence Detection System Applera Corporation Quantifiler™ Human DNA Quantification Kit Applera Corporation Quantifiler™ Y Human Male Quantification Kit Qiagen Qiamp DNA Blood Mini Kit Thomas D. Holland, PhD - W23 JPAC Central Identification Laboratory (Employee) 4 Bart Hoogeboom, MS - C18 3D Studio Max Software Jurian A. Hoogewerff, PhD - W10 NITECRIME (European Union Funded Network) (Grant/Research Support) Agilent New Wave (Demonstration Equipment) Katie M. Horsman, MS - B137 Applera Corporation Quantifiler™ Applera Corporation, Inc. Quantifiler™ Y Applied Biosystems, Inc. Primer Express® Applied Biosystems, Inc. Quantiblot™ Kyle A. Hoskins, BS - B102 Clorox® Bleach Fluorescein (provided by Gabor Patonay/Georgia State University) JOY® Dishwashing Detergent Max M. Houck, MA - B59 NIJ (Grant/Research Support) – B59 Prentice Hall/Pearson Education (Other Financial/Material Support) – W3 Rosalyn G. Huss, BS, RN - C49 Legal Nurse Services (provided by Center for Toxicology and Environmental Health, LLC) I Alice R. Isenberg, BA, MS, PhD - B132 Federal Bureau of Investigation Laboratory (Employee) J Jack L. Jacobia - D32 National Institute of Justice (Grant/Research Support) Richard L. Jantz, PhD - W20 FORDISC (provided by the University of Tennessee) SAS (provided by the University of Tennessee) SYSTAT (provided by the University of Tennessee) Cassie L. Johnson, MS - B99 Applied Biosystems, Inc. Y-Filer Promega PowerPlex® Y Kristen Johnson, BS - B108 Cedar Crest College (Grant/Research Support) Promega SE33 Kit Kimberley A. Johnston, MSc Government of Ontario (Employee) – B114, E19 Roger W. Jones, PhD - J6 National Institute of Justice (Grant/Research Support) Agilent Technologies HPLC pumps Agilent Technologies HPLC column Digilab Finnigan FT-IR Spectrometer HPLC/FT-IR Interface Lab Connections Mass Spectrometer Micro-Tech Susan W. Jones, PhD, MFS - B69 Department of Defense (Grant/Research Support) Applied Biosystems, Inc., Inc, Taqman® Idaho Technologies, Inc Thermocyclers MJ Research, Inc. Chelsey A. Juarez, BA - H37 University of California at Santa Cruz, UCSC Chicano Latino Resource Center (Grant/Research Support) Jane S. Juusola, BS - B172 Federal Bureau of Investigation (Grant/Research Support) * Presenting Author K Michelle Kaye, MA - H44 Forensic Sciences Foundation Lucas Research Grant (Grant/Research Support) Janine A. Kelly, MSc - D38 National Research Foundation (NRF), South Africa (Grant/Research Support) Ronald L. Kelly, BS - W16 3SI Security Systems FlexPac™ 3SI Security Systems Octapus™ 3SI Security Systems Scorpion®/Currency Guard United States Currency Protection Corporation Julie B. Kempton, MS, MFS - B167 Prince George’s County Police Department (Employee) Nam Yee Kim, PhD - K8, K12 National Institute of Scientific Investigation (Speaker’s Bureau) Rebecca J. Kirby - G68 Ferris Faculty Grant (Grant/Research Support) Walter E. Klippel, PhD - H1 Department of Justice Law Enforcement Innovation Center (Grant/Research Support) Eva E. Klonowski, PhD - H75 International Commission on Missing Persons - ICMP (Employee) Katherine A. Koen - D50 Federal Bureau of Investigations (Grant/Research Support) Jason C. Kolowski, MS - B130 Fisher Scientific LabCorp, Co. Mideo Systems, Inc. Roche Applied Science Rijad Konjhodzic, BS - B8 International Commission on Missing Persons - ICMP (Employee) Applera Corporation Quantifiler™ Promega PowerPlex®16 Dave Kontny - SS2 Florida International University (Other Financial/Material Support) Robert D. Koons, PhD - W10 NITECRIME (European Union Funded Network) (Grant/Research Support) Agilent New Wave (Demonstration Equipment) Benjamin E. Krenke, MS - B139 Promega (Employee) L Marrah E. Lachowicz, MFS - SS2 Florida International University (Other Financial/Material Support) Amrita Lal-Paterson, MSFS - B76 ReliaGene Technologies, Inc. (Employee) Qiagen EZ1, M48 Christopher Latkoczy, PhD - W10 NITECRIME (European Union Funded Network) (Grant/Research Support) Agilent New Wave (Demonstration Equipment) Marc A. LeBeau, MS - W16 3SI Security Systems FlexPac™ 3SI Security Systems Octapus™ 3SI Security Systems Scorpion®/Currency Guard United States Currency Protection Corporation Benoît Leclair, PhD - B110 Applied Biosystems, Inc. ChargeSwitch™ Applera Corporation Quantifiler™ * Presenting Author Cheng-Lung Lee, MFS - B115 Applied Biosystems, Inc. AFLP Plant Mapping Kit Qiagen, Inc.Plant DNeasy Extraction Kit Mark D. Leney, PhD - B129 Joint POW/MIA Accounting Command (Employee) Chris Lennard, PhD ChemImage Condor Chemical Imaging System – B25, B148 Digilab FTIR Imaging Sprectrometer with a 64x64 MCT Detector – B29 Polilight VCS-2000 Foster & Freeman (VCS2000) – B25 Dan Levitt, DDS - F18 WinID (provided by the OCME City of New York) Adobe® Photoshop® (provided by the OCME City of New York) Linda A. Lewis, PhD - W3 Prentice Hall/Pearson Education (Other Financial/Material Support) Jih-Heng Li, Dr - B52 National Bureau of Controlled Drugs Department of Health (Grant/Research Support) Ling Li, MD - G71 NICHD (Grant/Research Support) Richard Li, PhD - B5 Sam Houston State University (Grant/Research Support) Eng Seng Simon Lim, BS - B3 Health Sciences Authority - Singapore (Employee) Applied Biosystems, Inc. Identifiler™ Applied Biosystems, Inc. Genotyper™ Promega Powerplex® 16 Whatman®, FTA™ Collection Card Wen-Ling Lin, PhD - G70 IFM 91-T06 (Grant/Research Support) Laura L. Liptai, PhD - W7 Stark rxp (Other Financial/Material Support) Amanda L. Lokar, BA, BS - D4 New Orleans Police Department Scientific Criminal Investigation Division (Other Financial/Material Support) Nikon 5700 (provided by New Orleans Police Department Scientific Criminal Investigation Division) Pentax K1000 (provided by New Orleans Police Department Scientific Criminal Investigation Division) Kodak 35mm Film (provided by New Orleans Police Department Scientific Criminal Investigation Division) Compact Flash Card (provided by New Orleans Police Department Scientific Criminal Investigation Division) Katherine E. Long, BS - B166 Extraction, Amplification, Electrophoresis, and Analysis of STR DNA (provided by Applied Biosystems, Inc. and Millipore) José A. Lorente, MD, PhD - B101 Whatman FTA® Paper Applied Biosystems, Inc. Identifiler Kit™ José A. Lorente, MD, PhD - B9 Whatman (Grant/Research Support) Whatman FTA® Paper M Vahid Majidi, PhD - SS2 Florida International University (Other Financial/Material Support) Joseph J. Maltese, JD, MJS - W7 Stark rxp (Other Financial/Material Support) Holland Maness, BS - F3 ToothPrints® (provided by Kerr Dentistry) Mary H. Manhein, MA - SS2 Florida International University (Other Financial/Material Support) Julia Martin, MD - F27 Adobe® Photoshop® 5 Paul Martin, PhD - B20 CRAIC Technologies (Employee) CRAIC Technologies UV-Visible-NIR Microspectrophotometers JoAnne Marzowski, PhD - B34 Alpharma USPD Inc. Triple Antibiotic Ointment Ansell Healthcare Products Inc. Lifestyle Condoms Ultra Sensitive With Spermacide Avon Products Inc. Skin So Soft Lotion Body Shop International Cocoa Butter Moisturizing Stick Clinique Laboratories Inc. Clinique Daily Face Protectant Dayton Hudson Corp. Target Allergy Cream Inter-American Products Inc. Hydrocortisone Cream Johnson and Johnson Consumer Products, Inc. K-Y Jelly Kendall Company Kendall Thermazene Loreal USA Inc. Loreal Future Moisturizer Neutrogena Corp. Neutrogena Oil Free Moisturizer Neutrogena Corp. Neutrogena Sunblock SPF 45 Schering Plough HealthCare Products Inc. A+D Original Ointment Schering Plough HealthCare Products Inc. Coppertone Sport Sunblock SPF 15 Schering Plough HealthCare Products Inc. Coppertone Sport Sunblock SPF 30 St. Ives Laboratories Inc. Extra Relief Advanced therapy Lotion Bruce R. McCord, BS, PhD - K28 National Institute of Justice (Grant/Research Support) Kirstin McDonald, BA, BS - B125 Alden Leeds, Inc., Maxibrom Alden Leeds, Inc., nu-clo® 7 Day Tablets Alden Leeds, Inc., nu-clo® Concentrated Granules Clorox Company Clorox® Automatic Bowl Cleaner Global Household Brands X-14 Anti-Bacterial Toilet Bowl Cleaner Housechem American Fare® Automatic Toilet Bowl Cleaner with Bleach Housechem RiteAid Automatic Toilet Bowl Cleaner with Bleach Rickett & Colman Sani-Flush® Bleach Puck Plus® Automatic Toilet Bowl Cleaner WD-40 Company 2000 Flushes® Bleach Automatic Bowl Cleaner Richard W. McLay, PhD, PE - W7 Stark rxp (Other Financial/Material Support) Sara McNorton, MS - B58 Atomobile Body Fillers Automobile Spot Fillers Terry Melton, PhD - B131 Mitotyping Technologies (Employee) James R. Millette, PhD - C42 Microscopical Analysis (provided by MVA Scientific Consultants) Lisa M. Misner, MS - B127 National Institute of Justice (Grant/Research Support) Andre A. Moenssens, JD, LLM - W7 Stark rxp (Other Financial/Material Support) Shirly Montero, PhD NITECRIME (European Union Funded Network) (Grant/Research Support) – W10 Agilent – W10 Foster & Freeman Grim II – B159 New Wave (Demonstration Equipment) – W10 New Wave UP-213 Laser Ablation System – B159 Perkin Elmer ICP DRC Plus – B159 Stephen L. Morgan, PhD - B151 Federal Bureau of Investigation (Grant/Research Support) Keith Morris, PhD - W3 Prentice Hall/Pearson Education (Other Financial/Material Support) Robert J. Morton, MS - D41 Federal Bureau of Investigation (Other Financial/Material Support) 6 Robert F. Mothershead II, MA - W16 3SI Security Systems FlexPac™ 3SI Security Systems Octapus™ 3SI Security Systems Scorpion®/Currency Guard United States Currency Protection Corporation Dawn M. Mulhern, PhD - W2 Leica Microsystems Microscopes (Demonstration Equipment) Stacey Murnyak, BS - B146 NYC OCME Department of Forensic Biology (Employee) Applied Biosystems, Inc. ABI Prism® 3100 Genetic Analyzer Applied Biosystems, Inc. Quantiblot Bio RAD Micro Bio-Spin Chromatography Columns Promega DNA IQ Promega PowerPlex®16 N Huma Nasir, BS - B140 Applera Corporation Quantifiler™Y Reliagene Technologies, Inc. Y-Plex™12 Adam Negrusz, PhD, DSc - W15 SPE Products & Techniques (Unlabeled/Investigational Use) Varian, Inc. (Employee) Varian, Inc. SPE Phases Kristin N. Nestor, BS, BS - B15 TSWG (Grant/Research Support) Alexander A. Nieuwland, PhD - B150 Federal Bureau of Investigation (Grant/Research Support) Beckman-Coulter Biomek 2000 Workstation Beckman-Coulter P/ACE MDQ Capillary Electrophoresis System Waters Micromass Q-Tof Micromass Spectrometer Henry C. Nipper, PhD - K5 Zeneca Pharmaceuticals Seroquel (Quetiapine Fumarate) H. Dale Nute, PhD - B47 Florida State University, Panama City (Employee) Metal Detector (Unlabeled Pharmaceutical/Investigational/ Medical Device) O William R. Oliver, MD - D27 Multiple Web Pages Databases Kerry L. Opel, MA, BS - B147 NIJ 2002-IJ-CX-K007 (Grant/Research Support) Corbett Research Rotor-Gene RG3000 Millipore Microcon® YM-30 Stephen D. Ousley, PhD University of Tennessee, Knoxville (Consultant) - H22 FORDISC (provided by the University of Tennessee) – H22, W20 SAS (provided by the University of Tennessee) – W20 SYSTAT (provided by the University of Tennessee) – W20 P Robert R. Paine, PhD - W2 Leica Microsystems Microscopes (Demonstration Equipment) Frank J. Pappas, DDS - F18 Adobe® Photoshop® (provided by the OCME City of New York) WinID (provided by the OCME City of New York) Robert F. Pastor, PhD - H35 The British Academy (Grant/Research Support) * Presenting Author Gabor Patonay, PhD - B102 Clorox® Bleach Fluorescein (provided by Gabor Patonay/Georgia State University) JOY® Dishwashing Detergent Jeanette M. Perr, BS - SS2 Florida International University (Other Financial/Material Support) Donn N. Peterson, MSME, PE - W7 Stark rxp (Other Financial/Material Support) Joseph L. Peterson, DCrim - B62 Bureau of Justice Statistics (Grant/Research Support) Jerome L. Podorski, BS - SS2 Florida International University (Other Financial/Material Support) Natasha H. Poe, BS - B82 Applera Corporation Quantifiler™ Human DNA Quantification Kits Applied Biosystems, Inc. ABI Prism® 3100 Genetic Analyzer Applied Biosystems, Inc., Inc. AmpFlSTR® Profiler Plus™ Applied Biosystems, Inc., Inc. AmpFlSTR® Coflier™ PCR Amplification Kits Applied Biosystems, Inc. GeneScan® Analysis Software Applied Biosystems, Inc. Genotyper® Analysis Sofware Applied Biosystems, Inc. Quantiblot® Human DNA Quantification Kits The Bode Technology Group, Inc. Buccal DNA Collector Fitzco, Inc. Omni Swab Collector Fitzco, Inc. Sampact™ Pur-Wrap® Sterile Dacron® Swab (provided by Fitzco, Inc.) Qiagen, Inc. QIAmp® DNA Mini Kit Whatman® Indicating FTA® Micro Card (provided by Fitzco, Inc.) Mark M. Pollitt, MS - D13 University of Tulsa (Consultant) Maja Popovic, PhD - B75 ReliaGene Technologies, Inc. (Employee) Applied Biosystems, Inc. ABI Prism® 3100 Genetic Analyzer Applied Biosystems, Inc. 9700 GeneAmp® PCR System Applied Biosystems, Inc. Genotyper® Qiagen BioRobot 8000 DNA Extraction System Qiagen Biorobot Liquid Handling System Wallace DBS Puncher (provided by Perkin Elmer) Klaus Poulsen, MD - G85 Siemens CT-Scanner Somatom Plus4 Chang En Pu, MS - D53 Investigation Bureau, Ministry of Justice, Taiwan (Speaker’s Bureau) Genotyping Results (provided by Applied Biosystems, Inc.) Ken Pye, PhD - W10 NITECRIME (European Union Funded Network) (Grant/Research Support) Agilent New Wave (Demonstration Equipment) Michael P. Rickenbach, PhD - W16 3SI Security Systems FlexPac™ 3SI Security Systems Octapus™ 3SI Security Systems Scorpion®/Currency Guard United States Currency Protection Corporation Bernd Rieger, Dipl.-Phys. - D28 Adobe® Photoshop® Microsoft® Windows® Lovelle M. Rimando - D49 USDE Grant #P217A030070. (Grant/Research Support) Applied Biosystems, Inc. 377 XL DNA Sequencer (provided by the University of Hawaii BLAST Search Engine (Unlabeled Pharmaceutical/Investigational/ Medical Device) GenBank (Unlabeled Pharmaceutical/Investigational/Medical Device) Carol J. Ritter, MS - B108 Cedar Crest College (Grant/Research Support) Promega SE33 Kit James M. Robertson, PhD - B12 Federal Government (Grant/Research Support) James M. Robertson, PhD - B104 Federal Government (Grant/Research Support) Bayer Actril® Susan J. Roe, MD - G18 Bush Foundation Medical Fellows Program (Grant/Research Support) G. Sue Rogers, MSFS - B111 National Institute of Justice (Grant/Research Support) Applied Biosystems, Inc. AmpFlSTR® Applied Biosystems, Inc. GeneScan® Applied Biosystems, Inc. Genotyper® Applied Biosystems, Inc. ABI Prism® Hitachi FMBIO® Promega PowerPlex® Douglas E. Rohde, MS - K39 Merck Merckoquant® Cyanide Test Ann H. Ross, PhD - H18 North Carolina State University Grant (Grant/Research Support) Linda C. Rourke, MSFS, MPhil - B168 Amersham Biosciences 500 GenomiPhi Kit Lenny Rudin, PhD - D16 Cognitch (Grant/Research Support) Investigator Services (provided by Cognitech) Susanna Rudy, RN, MSFS Labock Technologies, Inc. One-Way(TM) Bullet Resistant Glass – B112 Pyng Medical Corporation F.A.S.T. 1™ Intraosseous System – D2 Q S Thomas Quatieri, PhD - D33 United States Government TSWG Air Force contract F19628-00C-0002 (Grant/Research Support) Kenneth Saczalski, PhD - C11 Bruce R. Pfaff & Associates (Grant/Research Support) Butler, Wooten, Fryhofer, Daughtery, & Sullivan (Grant/Research Support) Douthit, Frets, Rouse & Gentile (Grant/Research Support) Lowe, Eklund, Wakefield & Mulvihill (Grant/Research Support) Stong & Associates (Grant/Research Support) Nermin Sarajlic, MD - H70 International Commission on Missing Persons (Employee) Vincent J. Sava, MA - W23 JPAC Central Identification Laboratory (Employee) Michael I. Schaffer, PhD - K31 Psychemedics Corporation (Employee) Hair Analysis (provided by Psychemedics Corporation) Heather J. Schafstall, MS - W3 Prentice Hall/Pearson Education (Other Financial/Material Support) R J. Graham Rankin, PhD - B116 National Institute of Justice (Grant/Research Support) Machelle A. Reid, MFS - J15 Hewlett-Packard Company Pamela C. Reynolds, BS - W16 3SI Security Systems FlexPac™ 3SI Security Systems Octapus™ 3SI Security Systems Scorpion®/Currency Guard United States Currency Protection Corporation * Presenting Author 7 Kenneth G. Schoenly, PhD - G50 National Institute of Justice (Grant/Research Support) Susan Scholl, MS - W3 Prentice Hall/Pearson Education (Other Financial/Material Support) Rebecca L. Schuler, BS ChemImage Corporation (Employee) – D23, D71 ChemImage Corporation CI Print Macroscopic Chemical Imaging System - D71 ChemImage Corporation Fiber Database – D23 Microtrace Corporation Fiber Database – D23 Jeffrey Schweitzer, PhD - B162 National Institute of Justice (Grant/Research Support) National Aeronautics and Space Administration (Grant/Research Support) David V. Scott, JD - W7 Stark rxp (Other Financial/Material Support) Ronald Scott - W16 3SI Security Systems (Employee) 3SI Security Systems FlexPac™ 3SI Security Systems Octapus™ 3SI Security Systems Scorpion®/Currency Guard United States Currency Protection Corporation Jennifer Sears - B108 Cedar Crest College (Grant/Research Support) Promega SE33 Kit James A. Sebestyen, BS - B65 Office of Chief Medical Examiner (Speaker's Bureau) Dacron® (provided by Office of the Chief Medical Examiner) Corbett Research Rotor-Gene™ 3000 (provided by Office of the Chief Medical Examiner) Beckman-Coulter Biomek 2000 (provided by Office of the Chief Medical Examiner) Microcon 100 (provided by Office of the Chief Medical Examiner) Ismail M. Sebetan, MD, PhD - B109 Applied Biosystems, Inc. ABI Prism® 310 Genetic Analyzer Applied Biosystems, Inc. AmpFlSTR® Profiler Plus® PCR Amplification Kit Applied Biosystems, Inc. GeneScan® Carl M. Selavka, PhD - B162 National Institute of Justice (Grant/Research Support) National Aeronautics and Space Administration (Grant/Research Support) Arijana Selmanovic, BS - B4 International Commission on Missing Persons - ICMP (Employee) Applera Corporation Quantifiler™ Human DNA Quantification Kits Applied Biosystems, Inc. ABI Prism® 310 DNA Sequencers Applied Biosystems, Inc. ABI Prism® 3100 DNA Sequencers Applied Biosystems, Inc. ABI 7000 Sequence Detection System Promega® PowerPlex®16 Roche Molecular Systems, Inc. Taq Gold® Heather J. Seubert, BS - B102 Clorox® Bleach Fluorescein (provided by Gabor Patonay/Georgia State University) JOY® Dishwashing Detergent Kimberley Sharpe, MSc - B74 Applied Biosystems, Inc. AmpFlSTR® Profiler Plus® Applied Biosystems, Inc. ABI Prism® 377 DNA Sequencer Applied Biosystems, Inc. Genescan® Analysis Applied Biosystems, Inc. Genotyper® Software Kai-Ping Shaw, MD, PhD - G70 IFM 91-T06 (Grant/Research Support) Claire E. Shepard, MS - SS2 Florida International University (Other Financial/Material Support) Robin Shick - B108 Cedar Crest College (Grant/Research Support) Promega SE33 Kit 8 Michael E. Sigman, PhD - B41 National Institute of Justice (Grant/Research Support) Michael E. Sigman, PhD - B49 National Institute of Justice (Grant/Research Support) Kelli Sikorski - W3 Prentice Hall/Pearson Education (Other Financial/Material Support) Ronald L. Singer, MS - SS2 Florida International University (Other Financial/Material Support) David E. Sipes, MS - B98 Tecan Genesis Robot Qiagen DNA Extraction Kit Lejla Smajlovic, BS - B6 International Commission on Missing Persons (Employee) Applera Corporation Quantifiler™ Human DNA Quantification Kit Applied Biosystems, Inc. ABI Prism® 3100 Genetic Analyzer Applied Biosystems, Inc. ABI Prism® 7000 Sequence Detection System Applied Biosystems, Inc. ABI GeneAmp® PCR System 9700 Promega PowerPlex® 16 Keri L. Smith - B13 San Jose State University College of Applied Sciences and Arts (Grant/Research Support) Sorenson Microcentrifuge Tubes Amanda C. Sozer, PhD - B77 Louisiana State Police Crime Laboratory (Grant/Research Support) Pur-Wraps® sterile Dacron® Polyster-Tipped Applicator (provided by Fitzco, Inc.) Non-Contact Buccal Swab Laser-Cutting Instrument (Unlabeled Pharmaceutical/Investigational/Medical Device) Henry A. Spiller, MS - K20 Pfizer Visine® M. Kate Spradley, MA - W20 FORDISC (provided by the University of Tennessee) SAS (provided by the University of Tennessee) SYSTAT (provided by the University of Tennessee) Nicole Stalter - D47 USDE P217A990159 (Grant/Research Support) Lateefah A. Stanford, BS - C38 National Science Foundation (Grant/Research Support) Amy R. Stefan, BS - B149 Federal Bureau of Investigation (Grant/Research Support) Beckman-Coulter Biomek 2000 Beckman-Coulter P/ACE MDQ Capillary Electrophoresis System Thora S. Steffensen, MD - B21 The Icelandic Ministry of Justice (Grant/Research Support) Robin T. Stoehr, BS - B153 National Institute of Justice Award 2001-RC-CX-K003 (Grant/Research Support) K. Alan Stormo, MD - G62 Quick2000 Biohazard Escape Hood Samuel D. Stout, PhD - W2 Leica Microsystems Microscopes (Demonstration Equipment) Amanda B. Sturdevant, BS - B94 ATF-NLC (Other Financial/Material Support) 3M Scotch-Hutchinson Tape Other Scotch Hutchinson Tape Products Dale A. Sutherland, CChem - B119 Activation Laboratories, Ltd. (Employee) Anjali R. Swienton, MFS, JD - D59 National Insitute of Justice (Grant/Research Support) Jennifer A. Synstelien, MA - H86 Department of Justice Law Enforcement Innovation Center (Grant/Research Support) * Presenting Author T W Sabrina C. Ta'ala, MA - H14 Joint POW/MIA Accounting Command Central ID Lab (JPAC/CIL) (Employee) Allan Tereba, PhD - B73 Promega (Employee) Promega Differential Extraction System Mark D. Timken, PhD - B135 National Institute of Justice (Grant/Research Support) Applied Biosystems, Inc. STR Genotyping Kits Mitochon Amp Kits Roche Applied Biosciences qPCR Quantification Kits Bridget M. Tincher, MSFS - B79 National Institute of Justice (Grant/Research Support) Applied Biosystems, Inc. ABI Prism® Applied Biosystems, Inc. AmpFlSTR® Applied Biosystems, Inc. GeneScan® Applied Biosystems, Inc. Genotyper® Hitachi FMBIO® Promega PowerPlex® Richard E. Tontarski, PhD - W3 Prentice Hall/Pearson Education (Other Financial/Material Support) Hugh Tuller, MA - H24 International Commission on Missing Persons - ICMP (Employee) Andrew J. Tyrrell, PhD - W23 JPAC Central Identification Laboratory (Employee) Timothy W. Waldeck, JD - W7 Stark rxp (Other Financial/Material Support) Margaret M. Wallace, PhD - B173 PSC-CUNY #66248-00 35 (Grant/Research Support) Heather A. Walsh-Haney, MA - H77 University of Florida (Grant/Research Support) Tani G. Watkins, BA - B102 Clorox® Bleach Fluorescein (provided by Gabor Patonay/Georgia State University) JOY® Dishwashing Detergent John Watling, PhD - W10 NITECRIME (European Union Funded Network) (Grant/Research Support) Agilent New Wave (Demonstration Equipment) Gabriel D. Watts, BA - J19 Federal Bureau of Investigation Laboratory (Speaker's Bureau) Matthew J. Weber, BSE - C2 Accident Reconstruction Computer Software Douglas White - D14 National Institute of Standards and Technology (Employee) Commercial Software Applications Freeware Computer Applications Richard Winegar, PhD - B64 Federal Bureau of Investigation (Grant/Research Support) Amicon® Microcon® YM-10 Centrifugal Filter Devices Amicon® Ultrafree®-CL Centrifugal Filter Devices 0.45 mm Amicon® Ultrafree®-CL Centrifugal Filter Devices 5.0 mm Bio-Rad iQ™ Supermix Bio-Rad iQ™ SYBR Green Supermix Copan Enviro PBS TL Swab EpiCentre® BuccalAmp™ DNA Extraction Kit Machery-Nagel Nucleospin® DNA Trace Kit Millipore Multiscreen™ PCR Cleanup Plate MoBio UltraClean™ Mega Prep Soil DNA Kit MoBio UltraClean™Soil DNA Kit NIST Standard Reference Material® Urban Particulate Matter Qiagen DNeasy® Tissue Kit Qiagen QiaQuick® PCR Purification Kit Pluronic®F-68 Matt C. Wood, BS - C47 3-D Dispersion Modeling (provided by CTEH) U Douglas H. Ubelaker, PhD - W2 Leica Microsystems Microscopes (Demonstration Equipment) Noelle J. Umback, PhD - B141 Office of Chief Medical Examiner (Employee) Sayuri Umpierrez, BS - B39 Foster & Freeman (Other Financial/Material Support) Foster & Freeman LIBS System New Wave Research Laser Ablation System Perkin Elmer SCIEX ICP-MS System V Gerard J.Q. van der Peijl, PhD - B37 Alunite (provided by Riaza Mine in Spain) Alunite (provided by Rodalquilar Mine in Spain) Chatwood Safe EDAX Eagle m-XRF Instrument Europa Scientific Geo 20-20 IRMS Instrument Botanical Services (provided by National Herbarium of the Netherlands) IRMS Analyses (provided by Iso-Analytical, Ltd.) Perkin Elmer ELAN 6100 DRC Plus ICPMS Instrument Perkin Elmer OPTIMA 3000 ICP AES Instrument Stojko Vidovic, PhD - B7 International Commission on Missing Persons - ICMP (Employee) Promega PowerPlex® Human Identification Kit William Vilensky, DO, RPh - G89 Reckitt Benckiser (Grant/Research Support) Jessica C. Voorhees, MSc - B105 Federal Bureau of Investigation (Grant/Research Support) * Presenting Author Z Kathryn Zimmerman - G5 Millersville University Student Research Grant (Grant/Research Support) Jerry A. Zweigenbaum, PhD - K27 Agilent Technologies, Inc. (Employee) Agilent Technologies, Inc. LC/MSD TOF 9 Special Sessions SS1 The Big Bad Wolf is Worse Than Ever: Exploitation and Abuse of the Elderly Carla M. Noziglia, MS*, 8513 Northwest 47 Street, Coral Springs, FL 33067-3403; Michele E. Kestler, MS, Los Angeles Police Department, 555 Ramirez Street, Space 270, Los Angeles, CA 90012; Kathy Bell, MS, RN*, Tulsa Police Department, 600 Civic Center, Tulsa, OK 74103; William L. Leaver, BS*, Los Angeles County Sheriff's Department, 7717 Golondrinas Street, Downey, CA 90242; Ashraf Mozyani, PharmD, PhD*, Harris County Medical Examiner's Office, 1885 Old Spanish Trail, Houston, TX 77054; Douglas H. Posey, Jr., MD*, JAJ Forensic Center, Harris County, Texas, 1885 Old Spanish Trail, Houston, TX 77054; and Joseph N. Soos, BS*, Gray Murders/Gray Crimes Project, 1981Powell Creek Court, Charlottesville, VA 22911 Upon completion of this session, the participant should be able to clearly identify exploitation and abuse of the elderly and understand the scientific tools available for more effective prosecution. This presentation will impact the forensic community and/or humanity by better arming participants to combat elder abuse and exploitation in investigation, evidence analysis and prosecution. The generation labeled by Tom Brokaw as 'the greatest generation' has survived the horrors of a world war, the holocaust, the great depression and the atomic bomb. But another horror looms as the unscrupulous target these vulnerable silver citizens with fraud, robbery and, at times, murder. This workshop will provide insight in the identification of elderly exploitation and abuse, and discuss scientific and investigative tools, and methods which are effective to use. Too often, crimes against the elderly are categorized as abuse when, in fact, they can be much more. The topics focus on how the justice system, social services and the medical community are missing homicides and other crimes against the elderly. This workshop will also demonstrate the potential unintended and intended toxic effects of common prescription and over the counter medication with emphasis on the pharmacokinetics and pharmacological changes in elderly. Detecting forged signatures on documents such as wills, trusts and deeds will be presented. The indicators and evaluation of abuse of elderly noted in the emergency room as a result of sexual assault. The findings at postmortem examination: contracture, shaken adult syndrome, dehydration and malnutrition will be shown. The symposium will conclude with at discussion of the investigation into the death of an 87-year-old woman who was murdered for her estate. Elder Abuse, Crimes Against Elderly, Elder Sexual Assault SS2 Young Forensic Scientists Forum: Forensic Science Outside The Box Claire E. Shepard, MS*, 416 East Ponce De Leon Avenue, Decatur, GA 30030; Sheila M. Estacio Dennis, BA*, Office of the Chief Medical Examiner, 520 First Avenue, New York, NY 10016; Karly Buras, MA, Graduate Student, LSU, 3911 South Post Oak, New Orleans, LA 70131; Allison M. Curran, BS, Florida International University, 201 Southwest 116th Avenue, Apartment 304, Pembroke Pines, FL 33025; Amanda K. Frohwein*, 1663 Main Street Road, State Center, IA 50247; Christopher M. Gojcz, BS*, Drug Enforcement Administration, Special Testing & Research Laboratory, 22624 Dulles Summit Court, Dulles, VA 20166; Marrah E. Lachowicz, MFS*, 1300 East Orange Street, Tempe, AZ 85281; Jeanette M. Perr, BS*, Florida International University, 201 Southwest 116th Avenue, Pembroke Pines, FL 33025; Robert E. Barsley, DDS, JD*, Louisiana State University, School of Dentistry, 1100 Florida Avenue, Box 512, New Orleans, LA 70119-2714; Adam C. Becnel, BS*, Louisiana State Police Crime Laboratory, 376 East Airport Road, Baton 10 Rouge, LA 70806; James J. DiSarno, MS*, Drug Enforcement Administration, Northeast Laboratory, 99 Tenth Avenue, Suite 721, New York, NY 10011; Kenneth G. Furton, PhD, Director, International Forensic Research Institute, Florida International University, University Park, Miami, FL 33199; Kenneth W. Goddard, MS*, National Fish and Wildlife Forensic Laboratory, 1490 East Main Street, Ashland, OR 97520; Dave Kontny*, Director, National Explosives Detection Canine Program, Department of Homeland Security, Law Enforcement - Aviation Operations, 15500 Laurel Ridge Road, Montclair, VA 22026; Vahid Majidi, PhD*, Department of Justice, 950 Pennsylvania Avenue, NW, Washington, DC 20530; Mary H. Manhein, MA*, Forensic Anthropology and Computer Enhancement Services Laboratory, Louisiana State University, Baton Rouge, LA 70803; Jerome L. Podorski, BS*, Drug Enforcement Administration, Mid-Atlantic Laboratory, 1440 McCormick Drive, Largo, MD 20774; and Ronald L. Singer, MS*, Tarrant County Medical Examiner's Office, 200 Feliks Gwozdz Place, Fort Worth, TX 76104 Following the completion of this special session, the participants should be aware of the diversity of careers in forensic science, should understand how to compete for the FSF Emerging Forensic Scientist Award, should recognize the motivation behind and the guidelines for accreditation of forensic science academic programs, and should be aware of the realities in the field of forensic science according to young forensic scientists and crime laboratory directors. A breakfast session and an evening session will allow participants to mingle with students and emerging scientists from across the nation and to establish relationships with members for mentoring and networking. While at the breakfast session, participants will also have a chance to learn about resumé building and are encouraged to bring their own resumés to be critiqued. While at the evening session participants will be able to view graduate student posters and to mingle with their peers in a comfortable environment. As its role in society evolves, the field of Forensic Science continues to grow and incorporate the skills and knowledge of many different experts and disciplines. This year's special program will bring to the forefront some emerging forensic science fields, and consists of presentations by established members of the forensic science community. Emphasis has been placed on areas such as: the processing of clandestine labs, forensic odontology, wildlife forensics, forensic anthropology, and the importance of forensic science publications. The session will also stress the role of forensic science in homeland security, and will serve to make emerging forensic scientists aware of the opportunities available to them. Complete with a discussion panel on "Forensic Science in Louisiana," the session will give students and emerging Forensic Scientists an open forum atmosphere in which to discuss the realities of working as a forensic scientist, and the qualifications necessary to pursue this exciting career. This program should appeal to individuals with a strong desire to enter the field of forensic science as well as those with a few years of experience within the field and looking to get ahead. The session aims to cover a wide range of emerging fields through presentations, discussions, and interactions with new and established members of the forensic community. This program will provide a well-rounded understanding of different areas within forensic science. The objectives of this year's special session are as follows: • To introduce emerging fields in forensic science • To provide an understanding of the Emerging Forensic Scientist Award • To introduce not only emerging forensic scientists but also prospective forensic scientists to established forensic scientists through interaction, discussions, and presentations • To provide an opportunity for an open-forum discussion about the opportunities for forensic science in Louisiana • To increase involvement within the Young Forensic Scientists Forum. Education, Young Forensic Scientist Forum, Careers * Presenting Author SD1 Accreditation of Forensic Science Academic Program Through the AAFS FEPAC José R. Almirall, PhD*, Florida International University, Department of Chemistry, International Forensic Research Institute, Miami, FL 33199; Charles "Chris" Tindall, PhD*, Metropolitan State College, Department of Chemistry, PO Box 173362, Camput Box 52, Denver, CO -8021; Betty J. Horton, RR2, Box 145, Tower Hill, IL 62571, Max M. Houck, MA*, West Virginia University, 886 Chestnut Ridge Road, Suite 309, PO Box 6216, Morgantown, WV 26506-6216; Susan Hart Johns, MA, Illinois State Police, Division of Forensic Services, 630 East Washington Street, Springfield, IL 62701-1304; Karen W. Kershenstein, PhD, 11842 Clara Way, Fairfax Station, VA 22039; Peter M. Marone, MS, Division of Forensic Science, 700 North 5th Street, Richmond, VA 23219; Carl Selavka, PhD*, Massachusetts State Police Crime Laboratory, 59 Horse Pond Road, Sudbury, MA 01776; and Jay A. Siegel, PhD*, Indiana University, Purdue University, Chemistry, School of Science, 402 North Blackford, LD 326 D, Indianapolis, IN 46202 After attending this presentation, attendees will understand the process of accreditation through the AAFS FEPAC mechanism and be able to participate in the process as a reviewer of academic programs. Academic programs will also learn about the process of accreditation from different perspectives. This presentation will impact the forensic community and/or humanity by providing participants with a better understanding of the accreditation process and to encourage academic institutions to participate in the accreditation process. We also expect to encourage practitioners to invest in the process by volunteering to act as site reviewers. Forensic Science education will ultimately benefit from a bona fide accreditation process. The mission of the FEPAC is to maintain and enhance the quality of forensic science education through a formal evaluation and recognition of college level academic programs. The primary function of the committee is to develop and maintain standards and administer an accreditation program that recognizes and distinguishes high quality undergraduate and graduate forensic science programs. Attendance to this session will assist academic institutions who offer undergraduate and graduate degree programs in forensic science to prepare for the AAFS accreditation process through the Forensic Science Education Programs Commission (FEPAC). The session will also assist future site reviewers (academic and practitioners) in the preparation for the on-site reviews of academic programs. Accreditation of Academic Programs in Forensic Science, FEPAC/AAFS, Special Session ES1 Complex Forensic Science Issues in Highly Controversial Cases Michael M. Baden, MD*, New York State Police, Building 22, State Campus, Albany, NY 12226; Henry Lee, PhD*, Division of Scientific Services, Connecticut Department of Public Safety, 278 Colony Street, Meriden, CT 06456; and Cyril H. Wecht, MD, JD*, Allegheny County, 542 Fourth Avenue, Pittsburgh, PA 15219 The goals of this presentation are to assist attendees with the ability to objectively review and analyze forensic scientific issues; and to provide a better understanding of potential pitfalls involved in complex civil and criminal trials. This presentation will impact the forensic community and/or humanity by increasing understanding of relationships among forensic scientists, law enforcement agencies and attorneys. There are varying degrees of exactitude associated with each of the acknowledged fields of forensic science. However, none of these areas of professional expertise constitutes an absolute science. Accordingly, conflicting * Presenting Author views, even to the point of seemingly antithetical conclusions and opinions, may be encountered in any particular civil or criminal lawsuit. For various reasons, the more multi-faceted, controversial, and nationally publicized a particular case may be, the more likely are there to be highly conflicting opinions from acknowledged medical and forensic scientific experts. While this may be a source of consternation and even intellectual concern to many people, there is nothing inherently inappropriate or professionally unethical in such a situation. Reasonable, intelligent, and highly experienced experts may arrive at different conclusions in any particular case. Of course, the fact that the adversarial system is the universally accepted method of litigation trial technique in the United States, undoubtedly serves to foment and exaggerate these divergent viewpoints. This discussion will be designed to illustrate and perhaps explain how serious and significant differences among various forensic science experts may arise in nationally prominent cases. Presentations will include factual analyses of several such matters of recent vintage, including the Scott Peterson, Kobe Bryant, Jayson Williams cases and several others. Different perspectives will be discussed in order to ensure objectivity and fairness. Forensic Scientific Analysis, Complex Litigation, Adversarial System ES2 Requirements for Investigations into Allegations of Serious Negligence or Misconduct in Forensic Science Laboratories Joseph P. Bono, MA, Office of Forensic Sciences, Drug Enforcement Administration, 2401 Jefferson Davis Highway Alexandria, VA 22301; Kenneth E. Melson, JD, United States Attorney's Office, 2100 Jamieson Avenue, Alexandria, VA 22314; Barry Scheck, Cardozo Law School, 55 Fifth Avenue New York, NY; and Peter Neufeld; Attorney at Law; 99 Hudson Street; New York, NY During the past few years there have been a number of accusations directed at forensic science laboratories. These accusations have involved claims of negligence, misconduct or technical/analytical mistakes in the analyses of physical evidence with the result being possible miscarriages of justice. To the credit of many laboratories, there is a practice of self-disclosure when evidence of such impropriety exists. In most cases, the problems are addressed, notifications are made, justice is served and the integrity of the laboratory stays intact. In other cases, that is not the outcome. Now the federal government has mandated formal state government oversight to resolve these accusations. The Paul Coverdell Forensic Sciences Improvement Grant Program (FSIGP) was expanded by the 108th Congress to require that a state, as a condition of receiving federal funding for crime laboratories, certify that a formal, governmental process is in place to conduct independent external investigations of its laboratories. These investigations are to occur when there are allegations of serious negligence or misconduct substantially affecting the integrity of the forensic results committed by employees or contractors of any forensic laboratory system, medical examiner's office, coroner's office, law enforcement storage facility, or medical facility in the state that receives a grant under the Program. This presentation will offer an open and full discussion of the requirement for external investigations, the type of government entities that will oversee the investigations, the parameters of the investigations, and the effect on other quality assurance program obligations. The panel will consist of representatives of the Innocence Project at Cardozo Law School, who participated in drafting the amendment's language, and of ASCLD/LAB. Negligence, Misconduct, Forensic Science Laboratories 11 Breakfast Seminars BS1 The One Drop Rule: Is It Time For Science Rather Than Law to Define Race in America? BS2 The Pseudo-Medical Examiner and the Determination of Mode of Death: A Case History Ingrid A. Gill, JD*, 4835 North Kenmore Street, Chicago, IL 60640 Emanuel Tanay, MD*, 2977 Philadelphia Drive, Ann Arbor, MI 48103 After attending this presentation, attendees will learn the history of Louisiana's race classification laws and their impact on people of mixed heritage today. The presenter will draw on her tri-racial heritage in America and her Certificate of Ancestry by DNA Print Genomics to discuss the sensitive topic of race in America. This presentation will impact the forensic community and/or humanity by demonstrating evolving DNA technologies that can change the social and legal landscape of America. However, is the legal community ready to take up the challenge? Since this nation's inception, you have been defined as black if you had any known African black ancestry. This became known as the "one drop rule," meaning that a single drop of black blood makes a person black. Some courts have referred to it as the "one black ancestor rule" or the "traceable amount rule.” Anthropologists have designated it as the "hypo-descent rule," meaning that racially mixed persons are assigned the status of the subordinate group. During the Jim Crow era, many southern states had race classification statutes. Louisiana's race classifications statute in 1970 defined anyone as black whose ancestry is more than one thirty second black. Sussie Phipps challenged this law in the suite filed as Jane Doe v. State of Louisiana. Despite her blond hair and blue eyes, Ms. Phipps' birth certificate indicated that she was "colored" which contradicted the white classification she had checked off on her passport application. Her relatives admitted in depositions that they considered themselves colored. In 1983, district court declared her legally black. The Louisiana State Fourth Circuit Court of Appeals upheld the district court's decision. The court ruled that a party could not change their racial designation or their parents: "That Appellants might today describe themselves as white does not prove error in a document which designates their parents as colored. Of course, if the parents designation as ‘colored’ cannot be disturbed, their descendants must be defined as black by the traceable amount rule." Although noting expert testimony to the effect that the race of an individual cannot be determined with scientific accuracy, the court said the law of racial designation is not based on science, that "individual race designations are purely social and cultural perceptions and the evidence conclusively proves those subjective perspectives were correctly recorded at the time the appellant's birth certificate was recorded." At the rehearing, the appellate court affirmed the necessity of designating race on the birth certificate for public health, affirmative action, and that equal protection had not been violated. The Louisiana Supreme Court in 1986 declined to review the lower courts decisions. In December of 1986, the U.S. Supreme Court dismissed the appeal for want of a substantial federal question. Consequently, the one drop rule still legally determines who is black in America. Today, racial classification laws are voluntary. However, race information is still used for affirmative action and other important programs. Science does exist now where an expert can testify to the effect of the race of an individual based on Ancestry Informative Markers. The question is whether the one drop rule is ripe for reconsideration? After attending this presentation, attendees will learn that mode of death, unlike cause of death, cannot be determined by examining the body of the deceased. They will gain the conviction that mode of death determination, just like cause of death, is a subject that requires academic credentials and clinical experience. This presentation will impact the forensic community and/or humanity by inspiring the forensic community to establish professional criteria for the position of medical examiner. Mrs. Yvette Sherman and Dr. Tanay hope to increase the empathy of the forensic community for the parents of a child who suffered untimely death as the result of his or her own behavior. A County executive appointed a fireman (medic) to be the interim medical examiner for Brown County, WI. Raymond Sherman, the Green Bay Packers coach, and his wife, Yvette Sherman, had a fourteen-yearold son, Raymond, Jr. The father found his son in the garage with a fatal self-inflicted gunshot wound to the head. The medical examiner, Mr. Klimek, subsequent to an autopsy, issued a press release declaring that the youngster's death was suicide. Police investigation, forensic psychiatric reconstruction by Dr. Tanay, and the evaluation by a forensic pathologist, concluded that it was an accident. Mr. Klimek continued to insist to news media that it was suicide. A subsequent hearing before a trial judge determined that mode of death was accident. Nevertheless, Mr. Klimek issued another press release indicating that young Raymond's death was suicide. Mr. Klimek's sixteen-year-old daughter wrote a letter to the local paper, which was published, praising her father's courage to resist powerful forces in his effort to uphold the truth and professionalism. The controversy has had a great emotional impact upon the Sherman family, particularly Mrs. Sherman, the mother. Retrospective diagnosis of the suicidal state of mind of someone who is deceased depends primarily upon information from those who have known the victim. Physical evidence and autopsy results may be consistent with suicide but are not diagnostic. Psychosocial information is essential for the making of a diagnosis of depressive illness, which is the underlying cause of suicide. The gathering and interpretation of relevant psychosocial data requires a skilled professional. Such information has to be gathered and interpreted by professionals. Mrs. Sherman will discuss the impact this controversy has had upon her family and the community of Green Bay. Louisiana Race Laws, Legal Issues, Ancestry Informative Marker (AIM) 12 Medical Examiner, Mode of Death, Suicide BS3 Follywood — Forensic Fakes or Real Cases: How Movies Manipulate Forensic Truth Haskell M. Pitluck, JD*, 573 Lake Avenue, Crystal Lake, IL 60014; Linda Kenney, JD*, 15 West 53 Street, Apartment 18 B, New York, NY 10019; James E. Starrs, LLM*, The George Washington University, 720 20th Street, NW, Washington, DC 20052 After attending this presentation, attendees will understand the foibles of Hollywood influence on public thinking. This presentation will impact the forensic community and/or humanity by presenting the deluge of forensic cases as epitomized by the movie studios to show to the audience that not only are movies are not real life, but you can’t always believe what you see on television or in the movies. * Presenting Author Having received the Oscars for the last three years, the production team of Pitluck, Kenney, and Starrs – no guest appearances – will premiere their new movie “Follywood: Forensic Fakes or Real Cases; How Movies Manipulate Forensic Truth.” This year’s breakfast will impart the audience with a deluge of forensic cases as epitomized by the movie studios. Is Oliver Stone’s JFK real or false? Does the “Texas Chainsaw Massacre” or “Silence of the Lambs” capture the demented mind of Ed Gein? Is the “Ghost of Mississippi” a tale or a truth? This presentation will also veer into a Follywood stage set and discuss the news media. Do courtroom scenes as commented on by news anchors depict fact or fiction? Is the news coverage of events in the Middle East truth or lies? The audience this year will see the gamut from comedy to drama to documentary/docudrama. The viewer will determine whether Follywood is a mockumentary or reality. Forensics, Reality, Manipulation BS4 Houston, We Have a Problem: Burying Medical Examiner Mistakes J.C. Upshaw Downs, MD*, Coastal Regional Laboratory, Georgia Bureau of Investigation, 925A Mohawk Street, Savannah, GA 31419 After attending this presentation, attendees will understand the history of medical examiner accreditation and certification and be familiar with the nature of practice standards in forensic pathology. This presentation will impact the forensic community and/or humanity by assisting attendees to recognize danger signs of poor quality in forensic exams, specifically regards those conducted by the medical examiner. By developing a better understanding of the accreditation and certification processes, practitioners will develop an enhanced appreciation for and understanding of continued quality assurance efforts. The public often perceives the forensic lab in general and the medical examiner (ME) in particular as infallible. Recent high profile cases have centered on the dangers inherent in such a false sense of confidence, particularly in homicide cases. In any human-based system, errors are inherent. In the realm of the forensic pathologist, mistakes may well go unrecognized, as the proof of error is literally buried with the body. This is all the more tragic in that one of the arguments for establishment of a medical examiner system is the presumed reduction of incorrect death certification by lay coroners, the system supplanted by the ME. In this era of “evidence-based medicine,” one might reasonably ask what standards exist in the field (a point of significant interest within the ME community at present) to ensure that errors are detected before the evidence is “consumed in testing.” What is the evidence that the pathologist got the right answer? Particularly in the current period of intense public interest in and enthusiasm for forensics as a whole, efforts to ensure adequate funding for the crime lab and medical examiner office are endangered by low-quality exams by unqualified and/or charlatan practitioners. Utilizing several case studies, some common and not-so-common errors will be presented. The management dilemma in approaching recognized errors and “the problem scientist” will be addressed, specifically regards the approach to minimizing future mistakes and maximizing good science, to the benefit of all concerned parties. Benefit: Attendees will understand how to recognize danger signs of poor quality in forensic exams, specifically regards those conducted by the medical examiner. By developing a better understanding of the accreditation and certification processes, practitioners will develop an enhanced appreciation for and understanding of continued quality assurance efforts. Accreditation, Certification, Errors * Presenting Author BS5 The Death of Sir Harry Oakes Ronald L. Singer, MS*, Tarrant County Medical Examiner’s Office, 200 Feliks Gwozdz Place, Fort Worth, TX 76104-4919; Richard C. Froede, MD*, 3930 N Placita de la Escarpa, Tucson, AZ 85750; Suzanne M. Froede, MA*, 3930 North Placita de la Escarpa, Tucson, AZ 85750 The goal of this presentation is to describe the elements of inadequate investigation; understand the components of team investigation determine evidence and its value; and show what inadequate investigation anc court room preparation can do to a case. This presentation will impact the forensic community and/or humanity by showing what inadequate investigation and court room preparation can do to a case. Murdered in bed in his home in Nassau, Bahamas between midnight and dawn on the morning of July 9, 1943, Sir Harry Oakes was 68 years old at that time ass well as being one of the richest men in the world. His son-in-law, Alfred de Marigny, was accused of the murder, but later acquitted. The Duke of Windsor, who was the Royal Governor of the Bahamas, played a part in covering up clues that would have led to the real murderers. Harold Christie, a sometimes business partner of Oakes and the Duke, also appeared to be involved in the murder. Oakes was shot four times in the head and the bed set on fire. The body was badly scorched. Incompetent and dishonest detectives from Miami were brought in by the Duke of Windsor. de Marigny was accused of the murder and made to stand trial. However, without any evidence the jury could not convict him. Alfred de Marigny did escape a sentence in the Bahamas, but was forced to leave the islands. He had a difficult life and several attempts were made on his life. He was eventually allowed to enter the United States and became a citizen in 1975. A number of books have been written about the murder: King of Fools (1988), Who Killed Sir Harry Oakes (1983), The Life & Death of Harry Oakes (1959), sometimes with conflicting information and interpretation. In the latest book, Alfred de Marigny has written the story of his life: A Conspiracy of Crowns. The book was co-authored with Mickey Herskowitz and published by Crown Publishers, Inc., in New York, in 1990. Not only does he describe his fascinating life before moving to the Bahamas, but also writes about the death of Sir Harry Oakes and of suppressed evidence and silenced witnesses. The information of the FBI and other American agencies about the characters involved add extra interest to the account. Scotland Yard was never called in on the case and the British government has refused to reopen the case in spite of new evidence that has surfaced. The facts and idea will be presented using narrative and slides. Sir Harry Oakes, Investigation, Acquittal BS6 Bacon, Eggs, and Arsenic Suzanne Bell, BS, MS, PhD*, Bennett Department of Chemistry, West Virginia University, 217 Clark Hall, Morgantown, WV 26505-6045 After attending this presentation, attendees will have an understanding of one important aspect of the history of forensic science. Arsenic has been called "inheritance powder" because of its nefarious uses. It is an ideal poison because it can be given in incremental small doses. The symptoms, when they appear, mimic those of infectious diseases that were rampant before the introduction of antibiotics. This is ironic given that arsenic preparations were used effectively as anti-microbial agents for thousands of years. Ancient Egyptians and 18th century Parisians valued the pale look that only an arsenic face cream could supply; Lewis and Clark packed along arsenic-based treat13 ments for diseases that resulted from their enthusiastic hands-on diplomacy along their journey. This non-technical talk will tell the story of arsenic and how it defined early forensic science and forensic chemistry from Roman times to the Lafarge case, the first time chemical analysis was accepted by a court of law. This ground-breaking case was also the first time the testimony of a forensic scientist was commissioned and accepted by a court. This case did not stop homicidal poisoning, but it did bring it to light and demonstrated the value of everything from postmortem toxicology to quality assurance practices. It also drove poisoners to try different brews ranging from antimony and atropine to thallium and zinc. But once the race was on, chemists and toxicologists kept pace and as a result, poison is no longer the murder weapon of choice, having been replaced by high velocity lead poisoning. As will be seen, arsenic still has a devoted, if small, following. One of the most recent cases involved a poisoned breakfast. History of Forensic Science, Toxicology, Chemistry BS7 Sniper: How Digital Evidence Can Help Track and Convict the Bad Guy Richard W. Vorder Bruegge, ScM, PhD*, Federal Bureau of Investigation, Forensic Audio, Video and Image Analysis Unit, Engineering Research Facility, Quantico, VA 22135; Dara Sewell, BS*, Federal Bureau of Investigation, Computer Analysis Response Team, Engineering Research Facility, Quantico, VA 22135; Kenneth W. Marr, BSEE, MS*, Federal Bureau of Investigation, Forensic Audio, Video and Image Analysis Unit, Engineering Research Facility, Quantico, VA 22135 After attending this presentation, attendees will understand how computer forensics, audio analysis, video analysis, and image analysis of digital evidence can be applied, including precautions, safeguards, and limitations when analyzing digital evidence Many existing forensic disciplines employ newly developed digital techniques to conduct forensic examinations. Therefore, this presentation will impact the forensic community and/or humanity by providing recognition of the sources of digital evidence and describing the use of proper safeguards when examining digital evidence are of great importance to the forensic examiner. Digital evidence is taking a more important role in law enforcement, especially in high visibility criminal prosecutions and in counterterrorism. Many existing forensic disciplines employ newly developed digital techniques to conduct forensic examinations. Digital evidence has been recognized as a forensic discipline by the American Society of Crime Laboratory Directors, Laboratory Accreditation Board (ASCLD/LAB). Therefore, recognition of sources of digital evidence and use of proper safeguards when examining digital evidence are of great importance to the forensic examiner. This seminar describes several digital evidence examinations which when combined can provide crucial investigative insights and compelling criminal evidence. A fictional sniper case is presented with emphasis on several digital evidence examinations. The investigative process can use digital evidence results to identify links and associations that might otherwise have been overlooked; including, stolen identities, bank account passwords, encrypted file passwords, hidden computer data, surveillance video analysis, audio analysis of recorded background noise, and image enhancement. Computer forensic examiners gather and analyze evidence from a suspect's computer hard drive and digital storage devices. Forensic 14 principles of write-protection, imaging of computer data, precautions and limitations will be reviewed. Computer data analysis may reveal hidden or encrypted files stored on hard drives. Detection, recovery, and analysis of these hidden files require extensive training and experience of the CART examiner. Audio analysis of a suspect's recorded telephone threat and other audio surveillance recordings, as well as digital audio files can provide significant investigative guidance. Some handheld digital recorders now interface with computers to download files of recorded conversations that in turn can be attached to email messages and distributed over the Internet. Header information in the digital audio files can reveal descriptors for file size and characteristics indicating possible hidden, embedded information in the files. Video and image analysis employ digital evidence techniques during examinations of a suspect's facial comparison and height determination. Digital surveillance systems have special limitations that must be recognized and considered. Numerous issues confront the digital evidence examiner, including digital versus film resolution, compression, and documenting image enhancement operations. SWGIT Guidelines for acquiring, processing, analyzing, and preserving digital images will be reviewed. Digital evidence examinations are crucial in today's counterterrorism society. A variety of resources exist for support when conducting digital evidence examinations. These include the Scientific Working Group on Digital Evidence (SWGDE) and the Scientific Working Group on Imaging Technologies (SWGIT), which have issued guidelines for digital evidence examiners. Digital Evidence, Computer Forensics, Audio/Video Analysis BS8 Tom Krauss Memorial Bite Mark Breakfast: Forensic Witnesses and Their Vital Role in Special Victims Prosecutions Yolanda L. Rudich, Esq*, Sex Crimes/Special Victims Bureau, Richmond County District Attorney, 130 Stuyvesant Place, Staten Island, NY 10301 Special victims cases are among the most difficult for a District Attorney’s office to prosecute. Sex crimes, child abuse, domestic violence and crimes against seniors often are committed in places that are isolated from any witnesses. The victims are among the most vulnerable, often presenting physical and emotional impediments to gathering the required proof beyond a reasonable doubt. Increasingly, prosecutions are dependent upon forensic evidence, either as independent proof of criminal culpability or as legally necessary corroboration of witness testimony. Moreover, the popularity of TV shows such as “CSI” has persuaded jurors that such evidence is available and should be presented for their consideration. Thus, the role of the forensic witness has become critical in successful prosecutions. If the evidence obtained by these witnesses is to be most effectively presented within the criminal justice system, both prosecutors and forensic experts must have a clear picture as to how such evidence should be gathered and reported. Additionally, as professionals esteemed beyond the laboratory, you can embark on proactive programs in partnership with members of the criminal justice system that will help prevent crime in your own communities. Forensic Witnesses, Prosecution, Special Victims * Presenting Author BS9 Mmmmmm....MANCHESTER (Murder, Manslaughter, Medicolegal Machinery, Myth, and Mayhem in Manchester, United Kingdom) John D. Rutherford, BSc, MB ChB, FRCP (Edin), FRCPath, DMJ*, Salford Quays Forensic Pathology, PO Box 378, Salford, Greater Manchester M50 3UX, United Kingdom This study concerns the ten year period from January 1993 to December 2002. Autopsy reports were examined for all the "suspicious" deaths in Manchester (population approximately 2.75 million), United Kingdom. The outcome of the medicolegal process was established by pursuing these cases through the court records system. Approximately 1500 fatal cases fulfilled contemporary criteria of "suspicious". Roughly one third of the cases initially deemed to be "suspicious" turned out to be the result of criminal activity and, therefore, classified as "unlawful killing" of one sort or another. The ratio of "unlawful killings" to the total "suspicious deaths" (year by year), the causes of death, and the ways in which the coroners and criminal courts classified them will be outlined in the presentation. Inconsistencies in the classification system will be highlighted, some cases being difficult to categorize, raising the question of whether new terminology should be introduced. Although the recorded causes of homicide are proportionally different for the USA and the UK (mainly related to differences in availability of firearms) the criminal justice system in both countries relies upon the adversarial model which has intrinsic advantages and disadvantages. However, the focus is not on a comparative international study but hinges around potential criminal cases being missed by the British system, which derives from a coronial structure dating back to the thirteenth century. A classic example of such missed criminal activity was that of Dr Harold Shipman, Britain's most prolific serial killer, who murdered in excess of 200 of his own patients before detection; there have been other, less high profile, individual cases. The initial classification of a death as "suspicious" or "non-suspicious" (with consequent detailed investigation or virtually no investigation) is a crucial point at which mistakes can be made and potential criminal cases missed. A different system might stand a better chance of picking up otherwise undiscovered homicides. Serious questions arise such as "who should decide what is a suspicious death?", "if the right people are not now making such decisions, then who should?", "how do we attain consistency in the definitions of cause and manner of death?" and "has the advent of high technology at the turn of the 21st century made any significant difference to the end result (i.e. the court outcome) of the medicolegal process?". In keeping with the AAFS 2005 theme, this study straddles the turn of the century and is particularly relevant because at this meeting there is a focus on 21st century technology to combat crime. This presentation underlines the need for 21st century organizational thinking to run parallel with 21st century forensic science in order to revise ways of approaching the "Death Management System" from the first call for help from the bereaved relative right through to the final outcome in court. BS10 The Investigation of the Kidnapping of Danielle van Dam: The Physical Evidence Perspective Jennifer Shen, San Diego Police Department Crime Laboratory, 1401 Broadway, MS 725, San Diego, CA 92101; Tanya DuLaney, BS*, San Diego Police Department Crime Laboratory, 1401 Broadway, MS 725, San Diego, CA 92101 The goal of this presentation is to illustrate using a highly publicized trial, the importance of evidence collection, analysis, and presentation in court. In particular, this presentation highlights the importance of trace evidence in establishing contact timelines between the suspect and victim in this case. Due to the fact that the entire trial was televised we were able to, after the fact, select important points to emphasize in our presentation and demonstrate the types of direct and cross-examination a trace evidence analyst can expect in a trial of this magnitude. The issues of contamination, fiber and hair transfer, and significance of associations were addressed and are important and relevant topics. In the morning hours of February 2, 2002, a little girl was discovered missing from her bed. Thus began a massive search and a herculean effort by law enforcement agencies and citizens all over San Diego to find Danielle and bring her home. When her neighbor, David Westerfield, was identified as a suspect and her body was found, a series of events began to unfold, involving the San Diego Police Department and its crime laboratory in an investigation with time constraints and scrutiny unlike anything it had ever experienced. Crime laboratory personnel responded to scenes at the van Dam home, David Westerfield’s home, his motor home, his SUV, and the body recovery site. Westerfield was interviewed and polygraphed. Hundreds of pieces of evidence were collected, itemized, and analyzed. Suspicious behaviors, an unreasonable alibi, child pornography, and a failed polygraph, combined with Danielle’s blood on Westerfield’s jacket and in his motor home, gave the police sufficient cause to arrest him for the kidnapping and murder of Danielle van Dam. As the preliminary hearing approached, laboratory personnel worked feverishly to find more evidence. Latent prints located in the motor home above the bed were identified as Danielle’s. The blood and fingerprint evidence were presented at the preliminary hearing and Westerfield was bound over for trial. As the DA decided to seek the death penalty, the Westerfield defense insisted on a speedy trial, giving the crime lab only a few months to search through mountains of evidence. Danielle’s hair, her dog’s hair, carpet fibers, and clothing fibers began to emerge from the analyses. It then became necessary to prove that the evidence portrayed a recency of contact between the victim and suspect, and that the evidence was not transferred in an innocent fashion. In the end, after thousands of man-hours, involvement by nearly every laboratory section, and a grueling time schedule, David Westerfield was convicted of kidnapping and murdering Danielle and was sentenced to death. Kidnapping, Trace Evidence, Court Testimony Death Investigation, Homicide, Technological Advance * Presenting Author 15 BS11 On The Track of La Pérouse or the Unknown Man of Vanikoro Yves Schuliar, MD*, Institut de Recherche Criminelle de la Gendarmerie Nationale, 1, Boulevard Theophile Sueur, Rosny-Sous-Bois, 93110, France; Jean-Noel Vignal, PhD, Ircgn, 1 Boulevard Theophile Sueur, Rosny-Sous-Bois, 93110, France After attending this presentation, attendees will understand the role of the forensic sciences to help to find, to examine and to identify the skeleton of an individual who participated to a famous historical naval expedition in 1784. This presentation will impact the forensic community and/or humanity by demonstrating the interest of the cooperation between archeologists and a medico-legal team for the identification of historical cases. In 1784, Louis XVI, King of France, known as the “Sailor King” decided to organise an expedition to seach for the Northwest Passage from the Pacific side and to explore along the coasts of America, China, and Siberia and in the South Seas. He assigned a French navy veteran of the American War for Independence, the Comte Jean-François de Galaup de La Pérouse to undertake the expedition. Leading two ships, “La Boussole” and “L’astrolabe”, Jean-François de Galaup de La Pérouse initially explored the Russian and the Japanese coasts. Then the ships eventually left Australian shores on the 10th of March 1788. At that time, crews did not give any sign of life. Two hundreds sailors, scientists and officers of the King vanished. After several years of research, both shipwrecks were found on the VANIKORO Island. In November 2003, the Neo-Caledonian association “Salomon” found out, during an expedition in a coral reef, many ship parts among with an almost complete skeleton. After specifying its discovery conditions (a film will present the recovery expedition), authors describe the work carried out by a multidisciplinary team which studied the skeleton. The following points are discussed : • anthropological study • thaphonomical study • osteopathological study • post mortem delay assessment • odontological study • facial reconstruction • DNA profile analysis Hypotheses about the body’s identity are expressed, with respect to the expedition historical data. Submarine Archeology, Forensic Anthropology, Facial Reconstruction 16 * Presenting Author Luncheon Seminars L1 Frank Davis Cooks Naturally N’Awlins Frank Davis, WWL-TV Channel 4 Office, 1024 North Rampart Street, New Orleans, LA 70116-2487 From the heart of the historic French Quarter, from deep in the bayou country of southeast Louisiana, AAFS is proud to present Frank Davis, the absolute authority on New Orleans, Cajun and Creole cooking and one of the city’s most recognizable television chefs, cookbook authors, and gourmets. Born and raised in the heart of the Crescent City, Frank Davis began cooking New Orleans food at eight years old and hasn’t stopped since. Working with a number of notable chefs, Frank developed a unique culinary style of his own that can only be described as “Strictly N’Awlins.” In 1976, Frank Davis brought the Strictly N’Awlins style to WWL Radio for its first “live” two-hour-call-in cooking show. After five years on the air, he moved from radio to WWL TV and began airing “In The Kitchen with Frank Davis” in the two-hour morning news show. It’s still attracting an immeasurable morning audience and pulling unbelievable rating points. Frank wrote his first cookbook, “The Frank Davis Seafood Notebook,” in 1983. His second one, “Frank Davis Cooks Naturally N’Awlins,” came along in 1990. They both achieved “best-seller” status rapidly. His latest creation is “Frank Davis Cooks Cajun, Creole, and Crescent City.” It, too, is selling like the proverbial hotcakes all over the country! He is presently working on his next book, “Frank Davis Cooks N’Awlins Homestyle.” Plans are to have it ready sometime later this year. Cooking, Frank Davis L3 Justice From Both Sides of the Courtroom: From Homicide Detective to Criminal Defense Attorney Karen F. Ross, MD, Jefferson Parish Forensic Center, Harvey, LA 70058; Edward J. Rantz, JD*, Rantz and Associates, 1515 Poydras Street, New Orleans, LA 70112`; Karen F. Ross, MD*, Jefferson Parish Forensic Center, 2018 8th Street, Harvey, LA 70058; and Eric J. Hessler, JD*, 201 South Galvez Street, New Orleans, LA 70119 The pursuit of justice is a driving force in the lives of many in the various disciplines of forensic science. After attending this presentation, attendees will be introduced to two former homicide detectives for the New Orleans Police Department now attorneys whose practice includes criminal defense. This presentation will impact the forensic community and/or humanity by demonstrating the importance of the pursuit of justice from both the prosecution and defense viewpoints. These men have had the unique opportunity to participate in the criminal justice system on part of both the prosecution and the defense. They will relate their experiences as key fact witnesses for the state including some highly publicized cases such as that of a NOPD officer who was convicted of murdering a fellow officer and was sentenced to die. Both have also been involved in fatal shootings where they were forced to take the life of another person, so they may explain first hand what it is like to endure the scrutiny of criminal justice from that point as well. As criminal defense attorneys they continue to pursue justice, but from the other side of the courtroom, maintaining the position that all are entitled to an adequate defense and a trial before their peers as the system intends. Justice, Criminal Defense Attorney, Homicide Detective L2 What’s on the Menu Jason H. Byrd, PhD*, Office of the Medical Examiner, 1360 Indian Lake Road, Daytona Beach, FL 32124 After attending this presentation, attendees will gain an understanding of the Defect Action Levels allowed for food items. This presentation will impact the forensic community and/or humanity by providing the realization that insects are a common part of a well-balanced meal. Many societies have a social stigma against consuming insects as a food source. Our modern society may top the list of cultures having an aversion to “entomophagy”, the act of utilizing insects as food. However, our diet is more varied than we may imagine, and most of us unknowingly consume insects on a daily basis, as insects are a routine portion of our diet. Unfortunately, becoming aware of this simple fact may not allow you to avoid the inevitable. The United States Food and Drug Administration Center for Food Safety and Applied Nutrition establishes the maximum levels of natural or unavoidable defects in foods for human use that present no health hazard. Commercially processed egg, meat, and poultry products are regulated by the United States Department of Agriculture’s Food Safety and Inspection Service. Contaminants of fruits, vegetables, and other plants are regulated by the USDA’s Animal and Plant Health Inspection Service. Such levels are set because it is “economically impractical to grow, harvest, or process raw products that are totally free of non-hazardous, naturally occurring, or unavoidable defects”. This presentation focuses on the insect particulate matter present in the food we eat. Insects, Food, Defects * Presenting Author 17 Workshops & Workshorts W1 There is “Gold” in Mold: Forensic Evaluation and Litigation Issues in Mold and Indoor Air Pollutant Claims Mohan Nair, MD*, 5212 Katella, Suite 106, Los Alamitos, CA 90720; Robert Weinstock, MD*, 10966 Rochester Avenue, #4C, Los Angeles, CA 90024; Ajit S. Arora, MD, PhD*, UCLA, 8660 Woodley Avenue, North Hills, CA 91343; Ashok Jain, MD, MPH*, USC Medical School, 237 St. Albans, South Pasadena, CA 91030; and David Gyepes, JD, PhD*, 1990 South Bundy Drive, Suite 320, Los Angeles, CA 90025 After attending this presentation, participants will learn about the growing “toxic mold industry.” They will learn how to evaluate from medical, toxicologic and neuropsychiatric aspects of toxic mold claims. The connection between claims of mold related illness to other forms of abnormal illness behavior, and syndromes such as multiple chemical sensitivity, sick building /new building, fibromyalgia and chronic fatigue will be addressed. Mold spores are present in all indoor and outdoor environments and cannot be eliminated. Of more than 50,000 species of fungi, only about 150 are known to be human pathogens. While mold mycotoxins can cause mucosal irritation, there is no clear evidence of chronic, nonmucosal pathology in human beings, even in water-damaged buildings. Mold related litigation is described as “the next big thing” after asbestos. Between 1999 and 2001, there has been a thousand-fold increase in toxic mold-related insurance claims. A query on Google showed 114,000 hits for the term “Toxic Mold Neuropsychiatric.” These key words showed zero hits on Psych info and Medline. A third of the 600 million dollar homeowner claims paid out by Farmers Insurance in the state of Texas over the past two years was mold related. California became the first state in the nation to legislate moldrelated regulations, i.e., the Toxic Mold Protection Act, SB732, 2001. The magnitude of payouts have led to major insurance companies excluding coverage for mold related damage Though sometimes serious, physical illness in toxic mold claims is often of short duration. Attorneys and doctors with high profile toxic mold practices often emphasize n.europsychi.atric claims of disability and suffering from physical problems. This is especially true when there are a few robust findings on laboratory and physical examination. Allegations of brain damage are made on the basis of nonreplicable anecdotal and idiosyncratic interpretation of technologies, such as SPECT, PET and neuropsychological testing that often do not meet Daubert Standards. Body fluids are often sent for expensive and obscure tests. Findings of illness and disability may not be substantiated by face-to-face examination, the patient’s account of day-to-day functioning, on independent psychological testing, as well as by reviewing prior medical records and depositions. There is often evidence of pre-existing and concurrent factors, unrelated to the mold exposure in these individuals. Dr. Arora will cover key points in the medical examination. Dr Jain will discuss the toxicology of mold mycotoxins and factors in the physical and laboratory examination. Dr. Nair will present the steps in the psychological/psychiatric examination, and review of records Controversies in psychological/neuropsychological testing and neuroimaging findings will be discussed. Attorneys, toxicologists, occupational environmental medicine and mental health professionals who conduct Independent Medical and Psychiatric Examinations will benefit from this workshop. Mold, Indoor Air Pollutants 18 W2 Forensic Bone Histology Helen Cho, PhD*, Davidson College, PO Box 6934, Davidson, NC 28035-6934; Robert R. Paine, PhD*, Texas Tech University, Department of Sociology, Anthropology, and Social Work, Lubbock, TX 79409; Douglas H. Ubelaker, PhD*, and Dawn M. Mulhern, PhD*, National Museum of Natural History, Smithsonian Institution, Department of Anthropology, Washington, DC 20560; and Samuel D. Stout, PhD*, Ohio State University, Department of Anthropology, 124 West 17th Avenue, 244 Lord Hall, Columbus, OH 43210-1364 Upon completion of this workshop. the participant should have a better understanding of various applications of quantitative and qualitative histology of human skeletal tissue to aid in the identification of the unknown individuals. Much can be gleaned from skeletonized human remains, even if they are extremely fragmentary. This presentation will impact the forensic community and/or humanity by demonstrating how standard osteological methodologies coupled with specialized techniques. such as bone histology, can improve the likelihood that unknown skeletal remains will be identified. Histology, Bone, Microstructure W3 Educating Forensic Scientists for the 21st Century: Instilling the Forensic Mindset Suzanne Bell, PhD*, West Virginia University, Bennett Department of Chemistry, 217 Clark Hall, Morgantown, WV 26506-6045; Richard E. Tontarski, PhD*, ATF, Fire Research Laboratory, 6000 Ammendal Road, Beltsville, MD 20705; Sherry Brown, MS*, York College of Pennsylvania, Physical Sciences Department, York, PA 17405-7199; Aaron L. Brudenelle, BA, MS*, Tucson Police Department Crime Laboratory, 270 South Stone, Tucson, AZ 85701; Robert E. Gaensslen, PhD*, University of Illinois at Chicago, Forensic Science (M/C/866) College of Pharmacy, 833 South Wood Street, Chicago, IL 60612; Max M. Houck, MA*, Forensic Science Initiative, West Virginia University, 999 Chestnut Ridge Road, Morgantown, WV 26506; Linda A. Lewis, PhD*, Oak Ridge National Laboratory, 113 C Union Valley Road, Oak Ridge, TN 37830; Keith Morris, PhD*, c/o Suzanne Bell, PhD, West Virginia University, Bennett Department of Chemistry, 217 Clark Hall, Morganton, WV 26506-6045; Heather J. Schafstall, MS*, Oklahoma State Bureau of Investigation, 2132 NE 36th Street, Oklahoma City, OK 73111; Susan Scholl, MS*, New Mexico Department of Public Safety, PO Box 1628, Santa Fe, NM 87504-1628; and Kelli Sikorski*, c/o Suzanne Bell, West Virginia University, Chemistry/Forensic Science, 217 West Clark Hall, Morgantown, WV 26506-6045 The goal of this presentation is to assist attendees to design courses and course activities that serve the needs of students and the profession; understand the need for and foster the development, academic rigor, and acceptance; and define and understand the forensic mindset and use it to guide course and curriculum development. This presentation will impact the forensic community and/or humanity by assisting forensic educators in designing courses and curricula to meet the unique needs of forensic sciences as a career. It will also foster efforts to increase the academic rigor of forensic science with an emphasis on forensic chemistry. It will also provide a forum for practitioners and educators to discussion joint interest and opportunities. * Presenting Author The Technical Working Group on Education (TWGED) and the Forensic Science Education Programs Accreditation Committee (FEPAC) has done a laudable job of specifying the curricular requirements of a forensic science program. With the framework in place, attention is turning to execution. Educators, many of whom have not worked in the field, need guidance from the profession to flesh out the framework with meaningful courses, lectures, labs, and practicums. This need is particularly pressing in upper division and graduate-level courses where such materials are scarce to non-existent. However, with a need comes many opportunities. New analysts should leave university with a skill set unique to forensic science. This includes the ability to present meaningful testimony and reaching beyond the borders of their specialities. Instilling these skills, collectively the forensic mindset, is a challenge that forensic educators must rise to. This workshop will assist in this by defining and discussing the elements of the forensic mindset and offering practical ideas and suggestions for developing it in students. The primary goal of this workshop is to assist educators in fleshing out the framework of their programs and courses. Presenters will discuss and define the elements of the forensic mindset and offer practical guidance and suggestions for how to instill it within the context of an academic program. A second goal of the workshop is to discuss the need for academic acceptance of forensic science. Those who have crossed the great divide from forensic labs into the academic world are under pressure to increase the rigor of forensic science, particularly when it is incorporated into traditional disciplines and departments such as chemistry and biology. Many such departments regard forensic science/forensic chemistry/ forensic biology as little more that “blood spatter and crime scenes,” a misconception fueled by popular television shows and the flood of students generated. Overcoming this stereotype is vital for the field and can open new opportunities for research, funding, and collaboration. Courses need to be designed with this in mind. Conversely, successful courses and applications can capitalize on student interest and foster appreciation of science regardless of the student’s major or career plans. The morning session will present overviews and ideas from the perspective of students, educators, supervisors, and practitioners. The afternoon will delve into the nuts and bolts of course design, focusing on upper division courses. The discussions and applications have a chemistry flavor but the overriding principles and themes are broadly applicable. The target audience for this workshop are educators involved in or contemplating the launch of a forensic science program. Forensic supervisors,practitioners, and students with an interest in forensic education will also find much material of interest. Forensic Education, Professional Training, Continuing Education This presentation will impact the forensic community and/or humanity by serving to provide a foundation for forensic toxicologists and other interested parties so they may understand how and why agents can be toxic. It will marry the knowledge and observations of forensic pathologists with those of forensic toxicologists to aid in accurate Medical Examiner conclusions for cause and manner of death. Forensic Scientists evolve from a plethora of educational backgrounds that have not always provided a complete foundation for understanding the mechanism of toxicity by first comprehending how the body functions. This workshop can serve as a review, an introduction, or a continuing educational experience by offering an overview of anatomy, physiology, and pathology and an explanation of how each of these scientific fields are relevant to the basic knowledge of toxicity. The workshop will begin with a general description of anatomy. The speaker will highlight each of our body’s systems at the gross and cellular level. One-by-one anatomical systems such as the nervous, cardiovascular, musculoskeletal, gastrointestinal, respiratory, hepatic, renal, endocrine, and reproductive will be discussed. Once the body’s anatomical depiction is complete, the discussion will progress to the next topic of how toxins affect each of these systems. A forensic pathologist will explain how toxins adversely affect the body on a physiological and pharmacological level. Mechanisms of toxicity for specific anatomical regions will be discussed in detail including signs and symptoms, pathological findings, and pharmacological causes. For example, answers to questions such as: What exactly is pulmonary and cerebral edema and what causes them to occur when a CNS depressant is ingested in excess? What other pathological findings would be expected in cases involving pulmonary and cerebral edema? What other causes of death are consistent with these pathological findings and must be disproved before a pathologist rules the death toxicologically related? will be discussed. To further illustrate how toxins act on the body, specific agents will be discussed to explain how and why they kill or lead to serious and long-term injury. Considerations such as the route of administration, dosing history (e.g., acute vs. chronic, tolerance vs. non-tolerance) and drug-drug interactions all contribute to how and why a drug kills in one instance or individual but not another. These considerations are easier to explain once the foundations of anatomy, physiology, and pathology have been established. Finally, the panel of speakers will reconvene to address questions and further discuss topics of interest with the audience. At the conclusion of this workshop, the attendee will have a better understanding of how the body functions anatomically and physiologically, in addition to understanding how toxins act on and adversely effect the body. Anatomy of Toxicity, Physiology of Toxicity, Pathology of Toxicity W4 Anatomical, Pathological, and Physiological Foundations of Toxicity Diana Garside, PhD*, North Carolina Office of the Chief Medical Examiner, 1001 Brinkhous Bullitt Building CB#7580, Chapel Hills, NC 27599; Jeri Ropero-Miller, PhD*, North Carolina Office of the Chief Medical Examiner, 1001 Brinkhous Bullitt Building CB#7580, Chapel Hills, NC 27599; Michael J. Caplan, MD*, Department of Pathology and Laboratory Medicine, Division of Autopsy and Forensic Pathology, Medical University of South Carolina, Department of Pathology and Laboratory Medicine, 165 Ashley Avenue, Charleston, SC 29425; Alphonse Poklis, PhD*, Departments of Pathology, Pharmacology, Toxicology, Chemistry/Forensic Science, Virginia Commonwealth University, Medical College of Virginia, MCV Station Box 165, Richland, VA 23298; and Kenneth S. Snell, MD*, Mecklenburg County Medical Examiner’s Office, 618 North College Street, Charlotte, NC 28202 The goal of this presentation is to describe the anatomical systems of the body and how each functions; provide understanding of how toxins adversely affect the body and each of its anatomical systems; and discuss the common mechanisms, sign and symptoms, pathological findings, and actions of a toxin when it enters the body. * Presenting Author W5 Forensic Digital Evidence: Current and Prospective Scientific and Legal Issues Anjali R. Swienton, MFS, JD*, SciLawForensics, Ltd., 25 Walnutwood Court, Germantown, MD 20874; Gregory L. Hill, JD*, PO Box 451568, Kissimmee, FL 34745-1568; Susan W. Brenner, JD*, University of Dayton School of Law, Juseph E. Keller Hall, Room 421, Dayton, OH 45469-2772; Erin E. Keanneally, JD, MFS*, University of California, San Diego, San Diego Supercomputer Center, 9500 Gilman Drive, LaJolla, CA 92093-0505; Michael J. Menz, BS*, Hewlett Packard & Sacramento High Technology Crimes Task Force, 4510 Orange Grove Avenue, Sacramento, CA 95841; Fred C. Smith, JD*, Assistant United States Attorney, Federal Office Building, PO Box 607, Alburquerque, NM 87102; Michael C. Weil, MBA*, Huron Consulting Group, 550 West Van Buren, Chicago, IL 60607 After attending this presentation, attendees will have a basic understanding of some digital evidence investigatory strategies and examination procedures as well as evidence admissibility issues related to digital evidence 19 This presentation will impact the forensic community and/or humanity by opening discourse within the forensic community on timely issues related to accreditation of digital evidence laboratories, validation of examination techniques and trial strategies for trials involving various types of digital evidence. Currently, forensic digital evidence is anything but a science and is primarily an art, with few accepted techniques, standards of competence and performance and published data as to validity and reliability of results. Despite the increase in commercially available digital forensic tools and techniques, its introduction and use in the courtroom in criminal cases is largely limited to computer child pornography and Internet stalking cases. Digital forensic evidence is beginning to be introduced into the courtroom in murder, fraud, extortion, securities, and espionage cases. Meanwhile, digital forensics are employed in nearly every major white collar investigation and many aspects of complex civil litigation. A common theme that will begin to evolve as more and more cases are litigated, regardless of whether the case is criminal or civil, will be the attempt to develop accepted standards of reliability and validity to avoid Daubert challenges to digital evidence as a required routine in nearly every case. Challenges regarding the technology, ability of technological tools to access and recover such “evidence”, limitations or deficiencies in such tools to access and recover “evidence”, qualifications of the individual utilizing the tools and the ability of lay and expert witnesses to testify and effectively communicate in court about all of these matters will continue for the foreseeable future. Various interested sectors, including the network security application industry, have developed their particular niches and parameters for use of forensic tools and techniques. Digital forensics is beginning to be recognized as an area ripe for investigation and research. The workshop will place the rapidly changing and abstract landscape of litigation involving digital forensic evidence, into a more comprehensible and realistic picture and will be presented in four segments. The first segment will provide the attendee with an introduction to the field of forensic digital evidence, general terminology, widely used and tested tools, techniques, and their known limitations for the production and analysis of admissible forensic evidence. The second segment will be presented in the format of a criminal trial which will deal with several frequently encountered aspects of the problem of presenting admissible forensic digital evidence in the courtroom. Case facts derived from an actual case will be customized, with checklists and actual decision summaries to bring to life the concrete past examples and also provide guidance for handling future problems. A third segment will include a panelist discussion to assess the current status and prospective issues in forensic digital evidence. The panel includes: a prosecutor, a defense attorney, an academic legal scholar, a technical expert, an investigator, and an examiner. Each panelist will address certain issues relative to their area of expertise. Finally, a group discussion, with the attendees participating and presenting their own problems in advance to the panel for discussion and feedback will close out the session. At the conclusion of the workshop the participants should possess a basic understanding of the current status of forensic digital evidence and the scientific and legal issues that have been litigated to date. Through the panel presentations and group discussion, the forensic science community should be able to further explore the development of digital evidence to enhance its usage and reliability in the courtroom. W6 Serial Homicide, Myths, Legends and Facts Robert J. Morton, MS*, FBI NCAVC, FBI Academy, Quantico, VA 22135; James J. McNamara, MS*, FBI NCAVC, FBI Academy, Quantico, VA 22135; Mary Collins-Morton, BS*, FBI Washington Field Office, 601 4th Street, Northwest, Washington, DC 20535; and Gerard F. Downes, BS*, SSA - FBI NCAVC, FBI Academy, Quantico, VA 22135 The purpose of this workshop is provide investigators and medicolegal professionals with an understanding of serial murderers, their motives, methods of operation, victim selection, and body disposal scenarios highlighted through case examples. The focus of the workshop is on the practical issues involved in investigating and analyzing the actions of a serial murderer, the benefits of input from different disciplines and the need for cooperation between professionals. This presentation will impact the forensic community and/or humanity by providing participants a greater understanding of the “truth” about serial homicide, to include, the scope of serial murder, serial offenders and motivation, methods of operation, victim selection, body disposal, forensic techniques and collection strategies pertinent to serial homicide, and cooperative investigative strategies useful to successful case resolution. This workshop is targeted at providing investigators and medicolegal practitioners with a broad base of knowledge concerning serial murder, as well as a thorough understanding of the nature of serial offenders. Workshop will include a discussion of serial murder and it’s parameters, motivations of serial offenders, forensic issues and investigative issues. Workshop discussions will be augmented by numerous case examples, including several “high profile” cases. Serial homicide has long been an issue that generates much attention, from law enforcement, mental health practitioners, medicolegal professionals, and the media. There are a plethora of opinions. Law enforcement regard serial homicide as a painful anomaly, mental health practitioners look for causes, medico-legal professionals struggle with the results and the media inflates serial homicide into an epidemic. It’s no surprise that many myths and legends abound concerning serial killers. The FBI’s National Center for the Analysis of Violent Crime (NCAVC) is routinely consulted by federal, state and local authorities in a variety of cases of violent crimes, especially bizarre or serial homicides. The NCAVC has had extensive experience in assisting federal, state, and local law enforcement agencies in the investigation of serial homicides, and has reviewed hundreds of serial homicide cases for research purposes. Currently, the NCAVC is engaged in several research projects on serial offenders, including interviews of incarcerated serial offenders. The material presented in this workshop is based upon actual case experience, ongoing research, and current interviews with serial offenders. Serial Homicide, Serial Murderer, Victimology Digital Evidence, Technical Expert Testimony, Criminal Trial 20 * Presenting Author W7 Tutorial and Panel on Engineering Evidence and Lay Testimony Richard W. McLay, PhD, PE*, University of Iowa, 1231 Hamilton Court, Iowa City, IA 52245; Robert N. Anderson, PhD, PE*, RNA Consulting, Inc., 27820 Saddle Court, Los Altos Hills, CA 94022; Donald J. Anderson, BS*, Anderson Engineers, 13176 Pierce Street, Blain, MN 55434; Joe S. Cecil, PhD, JD*, Division of Research, Federal Judicial Center, One Columbus Circle, NE, Washington, DC 20002-8002; Laura L. Liptai, PhD*, Biomedical Forensics, 1660 School Street, Suite 103, Moraga, CA 94556; Joseph J. Maltese, JD, MJS*, New York Supreme Court, Home Port Court House, 355 Front Street, Staten Island, NY 10304; Andre A. Moenssens, JD, LLM*, The University of Missouri, 1760 East Poplar Road, Columbia City, IN 46725; Donn N. Peterson, MSME, PE*, Peterson Engineering, PO Box 47565, Plymouth, MN 55447-0565; David V. Scott, JD*, Scott, Forrest, and Bourne, PO Box 785, New Albany, IN 47151-0785; Thomas J. Vesper, JD*, Westmoreland, Vesper, Schwartz, and Quattrone, Bayport One, Suite 500, 8025 Black Horse Pike, West Atlantic City, NJ 08232; and Timothy W. Waldeck, JD*, Waldeck & Lind Law Firm, 1400 TCF Tower, 121 South 8th Street, Minneapolis, MN 55402 After attending this presentation, attendees will understand the three forms of engineering evidence and be able to evaluate lay engineering testimony and opinions. This presentation will impact the forensic community and/or humanity by demonstrating how a major problem with engineering testimony has been that laymen without proper training, using improper scientific methods, have been allowed to give engineering opinions in both criminal and civil cases. With the 1993 U.S. Supreme Court Decision on Daubert and the companion decision on Kumho Tire vs. Carmichael, methods have become available to challenge laymen giving questionable engineering testimony. The impact on our society and on humanity in general for this workshop will be to propose methods of fairness in evidence. This program, in two parts, presents: tutorials on engineering methods for use in accident reconstruction and a panel on lay engineering testimony. The tutorials are on technical content and precedents in the law: • Engineering fundamentals are presented for momentum and energy. • Biomechanics principles for accident reconstruction are illustrated. • Mapping in engineering analysis is shown to be crucial for records. • Simulation in vehicle impact and non-contact motion is presented. • Failure analysis of vehicle components is shown to be a factor in analysis. • Deposition methods for inquiring of lay engineering testimony are shown. • Daubert challenges are presented for engineering evidence. • Precedents in engineering evidence are presented. The panel discussion will focus on lay engineering testimony and the methods to be used to ensure that the evidence is good science and engineering. The speakers will be joined by the author of the amicus curiae brief for engineers, submitted to the U.S. Supreme Court in Kumho Tire vs. Carmichael; a Justice of the New York Supreme Court; and the Director of The Evidence Project at the Federal Judicial Center. A CD-ROM handout will include abstracts of the presentations. Engineering Evidence, Engineering Methods, Lay Testimony W8 Practical Homicide Investigation: Tactics, Procedures and Forensic Techniques Vernon J. Geberth, MS, MPS*, P.H.I. Investigative Consultants, Inc., PO Box 197, Garnerville, NY 10923; and Robert D. Keppel, PhD*, Sam Houston State University, Box 2296, Huntsville, TX 77341-2296 Upon completion of this workshop, the participants will have an understanding of the practice and theory of professional homicide investigation; they should better understand pertinent legal decisions, multiple crime scenes, components of a murder investigation, how time and distance as solvability factors affect murder cases, the management of the homicide investigations and the processing of the homicide crime scene. The instructors for this workshop have many years of homicide investigation experience. Currently, they both teach law enforcement officers from around the country about practical homicide investigation. At the Academy of Forensic Sciences, that same information can be passed on the forensic scientists from all disciplines. This presentation will impact the forensic community and/or humanity by providing forensic scientists with an understanding of the most modern techniques and procedures homicide investigators utilize to solve murder cases. Specifically, Commander Geberth will cover the practice and theories of professional homicide investigation involving equivocal death investigation, the homicide crime scene, the general and specific duties of the first officer, and duties at the crime scene for detectives. Dr. Keppel will focus on the empirical research on solvability factors in murder investigations. His presentation will show that the more information on the times and distances separating where the victim was last seen, the locations of the original contact between the victim and the killer, where the initial assault occurred, the murder site and the body site the more likely a murder case will be solved. The workshop also informs participants about managing the homicide case and the homicide crime scene process. This will include the use of blood enhancement reagents, the recognition and management of wound structures in shootings, stabbings, blunt force injuries and deaths by asphyxiation. The workshop concludes with a “who-done-it” and “who-is-it” rapemurder case presentation in which the victim was found in a wooded area with no identification and no immediate evidence leading to the killer. The participant will be taken through the entire investigative process from the finding of the body to the arrest of the suspect, using information about the site locations and the evidence at each site to identify the suspect and connect the suspect to the victim and, ultimately, resolve the case. Homicide, Murder, Solvability W9 Analysis of Samples From Clandestine Methamphetamine Laboratories Sanford A. Angelos, MSc, MEd*, and Camala L. Dubach, BSc*, U.S. Drug Enforcement Administration, 536 South Clark Street, Room 800, Chicago, IL 60605; David W. Love, BS*, U.S. Drug Enforcement Administration, South Central Laboratory, 10150 East Technology Boulevard, Dallas, TX 75220-4377; Nelson Santos, BA, MPA*, U.S. Drug Enforcement Administration, North Central Laboratory, 536 South Clark Street, Chicago, IL 60605, and Harry F. Skinner, MS*, U.S. Drug Enforcement Administration, Southwest Laboratory, 2815 Scott Street, Vista, CA 92081 The goal of this presentation is to present an overview of the predominant concerns surrounding the analysis of samples from clandestine methamphetamine laboratories. The attendees will receive information on the legal background and current status of methamphetamine as well as court testimony and production capacities of clandestine laboratories. This workshop will provide general information on current clandestine laboratory synthesis methods and specific protocols for the sampling and * Presenting Author 21 analysis of clandestine laboratory exhibits. Various reaction by-products and commonly identified adulterants found in samples and their significance will also be discussed. This presentation will impact the forensic community and/or humanity by demonstrating the ease of clandestinely manufacturing methamphetamine and the similarities in the varieties of chemical syntheses procedures, recognition and positive identification of the manufacturing process can present challenges. Currently, there are three major synthetic methods commonly used to convert ephedrine or pseudoephedrine to methamphetamine. The first is the hydriodic acid (HI) and red phosphorus (red P) method; the second is the iodine (I2) and red phosphorus method commonly referred to as the ‘Cold Cook’; and the third is the anhydrous ammonia (NH3) and either sodium (Na) or lithium (Li) metal method, known as the ‘Birch Reduction’ method. These methods have a regional character and can vary dramatically in production scale. All of these procedures require either ephedrine or pseudoephedrine as the precursor. Additionally, each of the procedures can have several variations. Given the ease of clandestinely manufacturing methamphetamine, and the similarities in the varieties of chemical synthesis procedures, recognition and positive identification of the manufacturing process can present challenges to forensic chemists and law enforcement personnel. The forensic chemist has two areas of concern in the analysis of clandestine methamphetamine laboratory samples. The first is the determination that there is a clandestine laboratory and that the material manufactured is in fact a controlled substance. The forensic chemist must be able to identify the synthesis procedure and the variations made by the operators. This requires a more systematic analytical approach. The second consideration is that the prosecution will usually require the forensic chemist to provide information that the site was a clandestine laboratory, and the clandestine laboratory’s production capability, especially if a substantial amount of a listed precursor is involved. Coupling the various complex and confusing legal aspects with the analytical difficulties, clandestine laboratory samples present the forensic chemist with significant challenges. The workshop will provide information to address the challenges stated above. An overview of the three current syntheses used to convert ephedrine or pseudoephedrine to methamphetamine encountered in clandestine laboratories will be provided. Various reaction by-products and commonly identified adulterants found in samples and their significance will also be discussed. In addition, the workshop will show how samples are identified to be submitted as exhibits and the procedures for taking samples at clandestine laboratories. Analysis protocols for liquids, both aqueous and organic, as well as solid samples, will be presented. Actual data will be provided. Clandestine Laboratories, Sample Analysis, Methamphetamine Synthesis W10 NITECRIME: Workshop on Trace Elements and Isotopes in Forensic Science Jose R. Almirall, PhD*, Department of Chemistry, International Forensic Research Institute, Florida International University, Miami, FL 33199; JoAnn Buscaglia, PhD*, FBI Laboratory, CITFSRU, FBI Academy, Quantico, VA 22135; Jim Ehrlinger, PhD*, University of Utah, Biology, ASB, Department of Biology, 257 South 1400 East, Salt Lake City, UT 84112; Jurian A. Hoogewerff, PhD*, Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA, United Kingdom; Robert D. Koons, PhD*, FBI Laboratory, CTFSRU, FBI Academy, Quantico, VA 22135; Christopher Latkoczy, PhD*, ETH Zurich, Lab of Inorganic Chemistry, HCI G111, Wolfgang-Pauli-Strasse 10, Zurich, 8093, Switzerland; Shirly Montero, PhD*, NFI, Netherlands Forensic Institute, Volmerlaan 17, Rijswijk, 2288 GD, The Netherlands; Ken Pye, PhD*, Prof. Kenneth Psy Associates, Ltd., Crowthorne Enterprise Centre, Crowthorne Business Estate, Old Wokingham Road, Crowthorne, RG45 6AW Berkshire, United Kingdom; and John Watling, PhD*, Curtin University of Technology, School of Chemistry, GPO Box U1987, Perth, 6845, Western Australia After attending this presentation, attendees will understand the basic principles of trace elemental and isotopic analysis, become aware of how these methods can be applied to forensic science and learn how to access these methods for their own work. This presentation will impact the forensic community and/or humanity by providing a better understanding of the application of these sophisticated methods to evidence analyses. Workshop participants will be exposed to a number of techniques and methods that can be immediately applied to improve the value of scientific evidence. The requirement to characterize and compare physical evidence from crime scenes, fraud and environmental casework is a major task in forensic science. Trace element and natural isotope profiles can assist in this process. New developments in instrumentation have created exciting possibilities for the routine “non-destructive” isotope and trace-element analysis of small and valuable specimens. Specialists from the NITECRIME Network (www.nitecrime.eu.com) will give an introduction on how to use trace elements and natural isotopic profiles to verify the authenticity and/or origin of raw materials, industrial products and materials, illegal drugs, foodstuffs, and human remains. Special attention will be given to the origin of trace element and isotopic variations in nature, the pro and cons of analytical systems and data presentation and interpretation. NITECRIME is an acronym for “Natural Isotopes and Trace Elements in CRIMinalistics and Environmental forensics”. It is an EU funded global network for developing and validating state of the art analytical methods and for disseminating these methods and practices to relevant users. Member organisations consist of forensic laboratories (FSS, FBI, BKA, NFI, US Customs and others) and academic institutions (Inst. of Food Research, Florida International University, ETHZ, Curtin Univ., Utah Univ., Otago Univ. and others) from around the world. (See also www.nitecrime.eu.com). The methods developed in the network are used to identify characteristic inorganic fingerprints not only from classical forensic materials like glass and bullets but also from food products, precious metals and human remains. Additionally the network is developing guidelines on how to interpret the multivariate data and how to present the results to a legal audience. Elemental Analysis, Isotope Ratio Analysis, Multi-Variante Data Analysis 22 * Presenting Author W11 Forensic Image and Video Processing Zeno J. Geradts, PhD*, Netherlands Forensic Institute, Laan van Ypenburg 6, Den Haag, 2497 GB, Netherlands; Richard W. Vorder Bruegge, PhD*, FBI Academy, Building 27985A, Quantico, VA 22135; Jurrien Bijhold, PhD*, Netherlands Forensic Institute, Volmeran 17, Rijswijk, 22884D, Netherlands; Bart Hoogeboom, MSc*, Netherlands Forensic Institute, Volmeran 17, Rijswijk, 2288, Netherlands; William R. Oliver, MD*, Georgia Bureau of Investigation, 533 Underwood Drive, Trion, GA 30753; Lenny Rudin, PhD*, Cognitech, 225 South Lake Avenue, Suite 601, Pasadena, CA 91101-3010; Carrie M. Whitcomb, MSFS*, National Center for Forensic Science, c/o University of Central Florida, PO Box 162367, Orlando, FL 32816-2367 After attending this presentation, attendees will undertand the possibilities are with digital images and video streams, and which techniques can be used in forensic science. Learn how quality assurance principles are applied to AV. During this workshop information will be provided on new developments of forensic investigation of (digital) images and video streams and the use of 3-dimensional computer modelling in forensic investigations. Traditional sources of images as evidence concern crime scene photography, and more specifically, photographs of fingerprints, tool marks, shoe prints and other impressions. A short overview of image processing techniques is given. Special attention is given to the introduction of artefacts by image processing (e.g. FFT on fingerprints). During the last 25 years the use of CCTV-camera systems has become widespread. Typical questions concern the quality and the selection of images from a specific camera in a multi-camera-recording. Digital processing of video streams for presentation and storage purposes, and the compression techniques that are applied in digital CCTV-systems, lead to questions about the integrity and authenticity of recordings. Also questions about image interpretation like facial recognition, body length, or car speed, often in low resolution, time lapse, or compressed images have increased. New sources of video streams and images are video recordings from handy cams, digital photo camera’s, internet and cellular phones. Typical questions about these recordings concern the integrity and authenticity of the recordings, the data compression techniques used, the synchronicity of sound and images, compensation for camera movement, and the conversion of a video stream to a higher resolution image. We will focus on methods for digital capture and analysis of analogue and digital multiplex surveillance recordings, state-of-the-art image enhancement techniques as contrast stretching and de-blurring, as well as new methods as super resolution, stabilising and automatic tracking. Since more images are being processed for forensic investigation, new methods have been developed for answering questions about the interpretation of images. Examples given: Is it possible to read a license plate number? Is our suspect, or his car, the one depicted in the image? What is the body length of the robber or the speed of a car? Is it possible to do a reconstruction of an accident or a shooting incident from the information in these images? Methods for image comparison, facial comparison with non standardized images, image reconstruction, and Photogrammetry are presented and discussed. Special attention is given to accuracy of the results and the impact on the conclusions from these investigations. Furthermore, there will be a hands on training during this workshop. Finally, some extra attention is given to the use of 3-dimensional computer modelling in forensic investigations, since we believe that these techniques will have an impact on traditional crime scene photography. Computer models and animations have been recently used for analysing video by superimposition of computer generated views of the model on the video images, for the visualisation of complex scenario’s in animations and for testing scenario’s against video footage and evidence in crime scene photographs. Examples: the reconstruction of car accidents from photographs, analysis of blood spatter patterns from photographs using a computer model of the crime scene, the visualisation of wound * Presenting Author channels in computer models of human bodies, the reconstruction of bullet trajectories, the reconstruction of a burglary using the limited information in dark images from a multi camera video recording, and the analysis of firework explosions from video recordings, photographs and geographical data. Special attention is given to modelling techniques, the accuracy of the models, methods for visualizing uncertainties and possibly erroneous suggestions coming from these visualizations. The use of image processing in the analysis of patterned injury of the skin, with emphasis on child abuse and as an aid in image analysis in forensic pathology will be discussed. The interpretation and recognition of image processing artifacts and image quality issues in forensic pathologic evaluation will be demonstrated. Image Processing, Video, 3D Reconstruction W12 The Pathological Examination of Deaths in the Elderly Gregory G. Davis, MD, MSPH*, Jefferson County Medical Examiner’s Office, 1515 Sixth Avenue South, Room 611, Birmingham, AL 352331601; Patricia J. McFeeley, MD*, Office of the Medical Investigator, MSC 11 6030, 1 University of New Mexico, Albuquerque, NM 871310001; and Richard E. Powers, MD*, University of Alabama at Birmingham and State of Alabama Department of Mental Health and Mental Retardation, 1530 Third Avenue South, SC961, Birmingham, AL 35294-0017 After attending this presentation, attendees will interpret clinical and pathological findings in the elderly; define clinical and pathological features of dementia and delirium; will be familiar with the incidence of abuse in the long term care setting; be able to interpret postmortem findings in residents of long term care settings, both anatomical and toxicological, in light of geriatric practice; and be able to distinguish nursing homes, assisted living facilities, home health, and hospice, including the regulation and expectations of these entities. As our society ages, questions concerning quality of care given to and received by elderly residents of long term care facilities, such as nursing homes or assisted living facilities, will grow more and more common. Often these questions will be directed to forensic pathologists, who are already familiar with injuries and death investigation. Nevertheless, elderly individuals, much like children, have their own special considerations when one is evaluating the state of their health. Geriatricians are familiar with these considerations, but forensic pathologists are less familiar. Moreover, forensic pathologists are less familiar with the long term care setting and the implications of various forms of care than are geriatricians. This presentation will impact the forensic community and/or humanity by presenting the forensic pathology of the elderly from a geriatrician’s point of view, thus enhancing the practice of forensic pathologists in serving the elderly. This workshop is designed for the pathologist who wishes to learn about the clinical manifestations and expected findings of advanced age in order to enhance the pathologist’s practice of forensic pathology. Because the setting in which death occurs is important in understanding how death occurs, the workshop will also cover the structure and governance of the various sorts of long term care facilities where many elderly individuals die. The workshop will consist largely of didactic presentations. In particular, the presentations on the clinical expectations of advanced aging and dementia and on long term care facilities will be given by a geriatric psychiatrist and neuropathologist who has served for years as the director of training for state nursing home accreditation inspectors. Handouts covering the topics will be provided, and there will be time for participant discussion and interaction. 23 One of the changes that may come with advanced age is dementia, the impairment of cognitive function that affects 47% of individuals who are 85 years of age or greater. As dementia progresses, the individual affected regresses so that, like a child, he is unable to care for himself. Significant behavioral abnormalities occur in up to 75% of demented persons. The stress that this condition places on the caregiver can lead to abuse which, like child abuse, is difficult to detect and difficult to prove with an amnestic, aphasic subject. At the same time, aging brings degenerative changes, such as osteoporosis or the friability of the skin and its superficial vessels, that can lead to findings that mimic abusive injuries. Careful correlation of the circumstances surrounding death with the findings at autopsy can establish or refute a claim of abuse, but this evaluation is made more accurately with knowledge of the structural and physiological changes that can be expected with advancing age. The circumstances surrounding death also depend on the environment in which death occurs. Knowledge of what constitutes reasonable and unreasonable care will help in the evaluation of a given death. Data collected by regulatory agencies can also provide valuable insights into past deficiencies in care identified during the survey process. Participants in this workshop will be taught clinical features of aging and dementia and the workings of various long term care facility organizations, such as nursing homes, assisted living facilities, and home health. Participants will have a better understanding of the special environment and medical considerations involved in deaths of elderly individuals from long term care facilities. Nursing Home, Assisted Living Facility, Elder W13 Human Factors, Performance, and Transportation Safety - The Rest of the Story: Beyond Alcohol and Other Drug Impairment Laurel J. Farrell, BA*, Colorado Bureau of Investigation, 690 Kipling Street, Suite 4000, Denver, CO 80215; Dennis J. Crouch, MBA*, Center for Human Toxicology, Biomedical Polymers Building, 20 South 2030 East, Room 490, Salt Lake City, UT 84112-9457; Bonnie M. Dobbs, PhD*, University of Alberta, Rehabilitation Research Centre, 3-62 Corbett Hall, Faculty of Rehab Medicine, Alberta, T6G 2G4, Canada; Jana M. Price, PhD*, National Transportation Safety Board, Research and Engineering (RE-10_, 490 L’Enfant Plaza E., Southwest, Washington, DC 20594; David L. Strayer, PhD*, University of Utah, Department of Psychology, 380 South 1530 East, Room 502, Salt Lake City, UT 84112-0251; Ginger S. Watson, PhD*, National Advanced Driving Simulator and Simulation Center, University of Iowa, 2401 Oakdale Boulevard, Iowa City, IA 52245; and John D. Weber, MD*, Webber and Associated, LLC, 12155 Southwest Wildwood Street, Portland, OR 97224-2919 The objectives of the workshop are to raise the awareness of toxicologists and other scientists who assess human factors contributions to crashes and highway safety to look beyond alcohol and drug related impairment; to be aware that cell phone and other distractions, fatigue, aging and medical conditions are important factors to consider in accident causation; and to gain an understanding of the models, methods and practical application of the data from behavioral performance studies. This workshop will have decided impact on the forensic community. Those associated with highway and other accident investigations will be introduced to a series of factors, outside of the usual investigative issues of alcohol and drug involvement, that play crucial roles in human performance. Historically, these factors have been largely overlooked by toxicologists, accident investigators and the general forensic community. Historically, toxicologists and other accident investigators have focused on the role of alcohol and other drugs as possible contributing factors to impaired performance. This focus has been guided in part by 24 epidemiological studies suggesting that alcohol and other drugs (marijuana, CNS stimulants and depressants) are over-represented in impaired, injured and fatally injured drivers. It has also been guided by controlled closedcourse and driving simulator studies demonstrating the impairing potential of many centrally acting drugs. However, highway and other accident investigations are far more complex than simply identifying a potentially impairing substance and assuming causation or even contribution. This workshop is designed to raise the awareness of toxicologists and other forensic investigators about a number of human factors (often over looked) that can affect performance in driving and other complex behavioral tasks. Data will be presented from an international panel of experts on human factors and performance in the areas of cell phone and other distractions, fatigue, aging, and medical conditions as well as models and methods for assessing impaired performance. As cell phone use increases, there have been growing concerns about the effects on highway safety and the constructs of effective legislation. Data will be presented comparing impairment from cell phone use to that of alcohol consumption. Fatigue has been shown to be a contributing factor in highway safety, yet it is often overlooked in accident investigations. The National Transportation Safety Board plays a leading role in this area of research and will discuss fatigue involvement in highway safety. Medical factors such as those affecting visual acuity, color discrimination, cardiac sufficiency, asthma, diabetes and psychiatric disorders and their potential contribution to driving will be discussed. Closely allied with the medical factors that may contribute to driver impairment are the complications and health issues associated with aging. Physicians already help their older patients preserve their driving abilities by providing health maintenance, but they also can play a more active role by assessing their patients for medical fitness to drive. Lastly, to fully understand the literature and research in the areas just discussed, one must have an understanding of driver distraction models, assessment methods, experimental measures and the applicability of the results. By understanding some of the research being done at the National Advanced Driving Simulator we will gain knowledge in all of these areas. From this workshop participants will gain an awareness of the potential contributions to accidents, crashes and highway safety of numerous human factors such as cell phone and other distraction distractions, fatigue, aging and medical conditions. Also, they will gain an understanding of the models, methods and practical applicability of the data from behavioral performance studies. Impairment, Human Factors, Safety W14 State of the Art Infrared and Ultraviolet Examinations of Documents by the Video Spectral Comparator Troy J. Eberhardt, BS*, U.S. Department of Homeland Security, U.S. Immigration and Customs Enforcement, 800 Westpark Drive, Suite 325, McLean, VA 22102; John L. Sang, MS*, Sang Forensic Document Laboratory, One Harbor Lane, Glen Head, NY 11545; Kristina Kovarik, BS*, Foster & Freeman, 46030 Nanekin Plaza, Suite 170, Sterling, VA 20166; and Gerald B. “Jerry” Richards, MS*, Richards Forensic Services, 15307 Alan Drive, Laurel, MD 20707 The goals of this workshop are to help the attendees understand the theory light relative to VSC Examinations; understand the theory and application of visible/infrared transmitted, incident, coaxial and oblique light examinations; understand the theory of longwave and shortwave ultraviolet incident and transmitted light examination; and understand and apply the appropriate filtration; and to provide understanding of basic microspectrometry applications (e.g., measure absorption, reflectance, transmission, and fluorescence spectra). This presentation will impact the forensic community and/or humanity by preparing the document examiner to better understand the theory behind the analysis and better explain his / her findings. * Presenting Author Infrared (IR), Ultraviolet (UV) and visible radiation has been used for many years in the examination of questioned documents revealing differences in inks, obliterations, erasures, alterations, deletions and insertions to documents. This workshop is going to explore the theory of light which makes these examinations possible. Jerry Richards will discuss the theory of these radiations. Kristina Kovarik of Foster and Freeman will explain the practical application of these theories and the latest technologies using a High Resolution Video Spectra Comparator. The workshop will cover incident visible, IR reflected, transmitted, and Luminescence examinations as well as short and long wave ultraviolet UV examination. Coaxial illumination and oblique visible and IR light techniques will be explored and demonstrated. Measurement of absorption, reflectance, transmission and fluorescence by the Microspectrometry will also be explained. The workshop will conclude with a group discussion lead by John Sang on the protocols for theses types of examinations. Infrared, Ultraviolet, Alterations W15 Practical Chromatographic Mechanisms Applied to Solid Phase Extraction Max B. Erwine, BS*, BGS Varian, 1634 Otte Avenue, Cincinnati, OH 45223; Adam Negrusz, PhD, DSc*, Department of Biopharmaceutical Sciences, College of Pharmacy, UIC, 833 South Wood Street, Chicago, IL 60612 After attending this presentation, attendees will have a working knowledge of Solid Phase Extraction, where it can be applied, a working knowledge of the fundamentals and a reference to trouble shooting and other resources. This presentation will impact the forensic community and/or humanity by improving bench knowledge of chromatography and the use of solid phase extraction. This half-day workshop will include a comprehensive discussion on all aspects of solid phase extraction techniques with some application examples. The discussion will be divided into Fundamentals, which will include reasons for using SPE, the steps of SPE, a chromatographic comparison of digital SPE vs. HPLC, silica and as a substrate for SPE, polar, non polar and ion exchange chemistries will be discussed in detail. Advanced and alternative approaches will detail improvements in the retention of polar compounds, trouble shooting, reduction of ion suppression in MS detectors, the use of polymers in SPE, packed bed and disk technologies and the use of solid supported liquid/liquid extraction techniques. Solid Phase Extraction, Chromatography, Sample Preparation W16 FBI Bank Security Device Workshop Marc A. LeBeau, MS*, and Ronald L. Kelly, BS*, FBI Laboratory, 2501 Investigation Parkway, Quantico, VA 22135; Joel Bartholf, BS*, 3SI Security Systems, Ashley Park, Suite 5A, 3902 Northside Drive, Macon, GA 31210; Phillip H. Grimm, BS*, U.S. Currency Protection Corporation, PO Box 6021, Scottsdale, AZ 85261; Robert F. Mothershead II, MA*, Pamela C. Reynolds, BS*, and Michael P. Rickenbach, PhD*, FBI Laboratory, 2501 Investigation Parkway, Quantico, VA 22135; and Ronald Scott*, 3SI Security Systems, 5891 Harmony Ranch, Aubrey, TX 76227 After attending this presentation, attendees will be able to further their knowledge in the area of bank security device products: past, present, and future. A validated procedure for the analysis of bank dye-pack chemicals will be introduced and available for scientists to implement. Various analytical techniques will be discussed so that current forensic examiners can tailor the procedure to what is applicable to their laboratory’s capabilities. * Presenting Author In order to adequately analyze bank security device evidence, a forensic scientist should possess much information about the dye-pack products and the procedure used to identify chemicals of interest. This presentation will impact the forensic community and/or humanity by attempting to provide examiners the knowledge and skills to not only analyze evidence related to bank security device dye-packs, but to adequately testify as an expert in this area. For many years the Federal Bureau of Investigation (FBI) has been the lead agency in the analysis of evidence from bank robberies. Bank security dye-packs expend chemicals when activated during a robbery and contaminate money and other pieces of evidence with specific chemicals. A forensic scientist should be kept up to date on products related to dye-packs as well as to analytical techniques and procedures used to detect the expended chemicals. There is a history in the evolution of dye-pack products, their manufacturers, and the analytical methods used to identify specific chemicals on evidentiary items. Active forensic scientists who are either currently working with bank dye-pack evidence or who are foreseeing a future in analyzing these types of cases will be offered pertinent information, a validated procedure, and opportunities to discuss issues with both company representatives and other qualified scientists. Case examples, evidence packaging, method development issues, and future products in the area of bank security devices will also be discussed. Bank Security Devices, Dye-Pack, Procedure W17 SWGIT Presents: Guidelines for Acquiring, Processing, Analyzing and Archiving Video and Image Data Richard W. Vorder Bruegge, PhD*, FBI, Forensic Audio, Video and Image Analysis Unit, Building 27958A, Quantico, VA 22135; Carl R. Kriigel, BS*, U.S. Army Criminal Investigation Laboratory, 4553 North 2nd Street, Forest Park, GA 30297; William R. Oliver, MD*, Georgia Bureau of Investigation, Northwest Regional Crime Laboratory, 533 Underwood Drive, Trion, GA 30753; and Mark Shuman, MD*, MiamiDade County Medical Examiner Department, Number 1 Bob Hope Road, Miami, FL 33136 Upon completion of this workshop, the participant will have a better understanding of how to incorporate proper procedures for handling image evidence into their law enforcement activities. They will know what sort of equipment and software they should utilize, how best to document their procedures, and will also know something about the legal; basis for the presentation of digital evidence in court. Images and video are intrinsic to law enforcement activities today. Crime scenes, suspects and evidence are photographed to document steps in an investigation. Surveillance images are seized and processed in order to reconstruct events and help criminals. Some images, such as latent print photographs, are processed and then analyzed to identify suspects. Many of these images ultimately find their way into the courtroom for use in prosecution. Analyses conducted on these images are frequently crucial to the successful completion of an investigation. The increased use of digital imaging technology in law enforcement has led many defense attorneys to raise multiple criticisms of it and law enforcement’s procedures, in an attempt to exclude such evidence. In this environment, it is more critical than ever that law enforcement take active measures to ensure that its images and imaging procedures can be defended in the court room. Recognizing this challenge, the FBI formed the Scientific Working Group on Imaging Technology (SWGIT) in 1997 to address the myriad of issues that can arise related to the science and technology of imaging. As of June 2004, SWGIT had formally published nine documents (with more in press) that address issues ranging from general guidance on chain of custody and the proper media for the preservation of image evidence, to procedures for image processing, the handling of video evidence, and advice on training in imaging. The purpose of this workshop is to acquaint law enforcement personnel with these documents and the 25 guidance contained therein, so that the attendees can incorporate these guidelines into the procedures within their own agencies and laboratories. Attendees will learn about chain of custody issues as they relate to crime scene photographs and surveillance video recordings from both analog and digital systems. They will also receive guidance regarding proper procedures for preserving such data and will learn where they can go to find out more about it. After a general overview, attendees will then be given some guidance regarding how to best utilize digital and traditional film systems for the acquisition of evidentiary photographs at crime scenes and in the laboratory. The selection of cameras and the steps necessary to acquire images sufficient for the purpose needed will be discussed. This will include a detailed explanation of image resolution and other factors that should be considered to determine which camera (and imaging system) is best suited for the purpose at hand. (It may surprise some to learn that digital imaging is not the answer to all of law enforcement’s imaging needs!) Next, attendees will learn about the variety of video surveillance systems in use throughout the country and SWGIT’s efforts to work with manufacturers to develop industry standards that meet the evidentiary needs of law enforcement. This section of the workshop will also address proper procedures for the capture and processing of video evidence by properly training personnel. Once images have been acquired in an investigation, they must be processed and analyzed to produce results meaningful to an investigation. SWGIT has developed guidelines regarding what sort of processing steps are most useful in a forensic setting and has also provided guidance on how to document those steps. Likewise, SWGIT is developing guidelines to provide imaging scientists with a set of “Best Practices”, to ensure that the testimony offered by imaging experts is supported by practices as rigorous as those applied in other forensic disciplines. Next, attendees will learn of the recent recognition of “Digital Evidence” as a discipline subject to accreditation by ASCLD/LAB. Included within this discipline is the subdiscipline of “Video and Image Analysis”. SWGIT recognizes that some confusion remains over what parts of a laboratory might be subject to accreditation under this discipline, and can provide guidance on this issue. Finally, many in law enforcement remain concerned regarding the admissibility of digital images within the courtroom. This workshop will provide attendees with a reference list of case law, as well as common sense approaches to this issue, that should help them ensure that their images, no matter their source, are admitted in court. Imaging, Image Processing, Digital Evidence W18 Shooting Reconstruction Timothy M. Palmbach, JD, MS*, University of New Haven, 300 Orange Avenue, New Haven, CT 06516; Robert K. O’Brien, MS*, State of Connecticut, Forensic Science Laboratory, 278 Colony Street, Meriden, CT 06451; Edward Jachimowicz, BS*, State of Connecticut, Forensic Science Laboratory, 278 Colony Street, Meriden, CT 06451; and Paul E. Kish, MS*, PO Box 814, Corning, NY 14830 The goal of this presentation is to discuss scene investigation and scientific principals involved in the investigation of a shooting incident. This presentation will impact the forensic community and/or humanity by demonstrating thorough investigation through scientific principals. This workshop will focus on the investigation and reconstruction of shooting incidents. Physical evidence and pattern interpretation at the shooting scene will provide valuable information in conducting a proper shooting reconstruction. In addition, laboratory analysis and interpretation of the physical evidence is critical to the reconstruction. Topics such as determination of bullet trajectory, glass interpretation, bloodstain pattern interpretation, GSR/Distance Determination and the national firearm database will be discussed. Numerous case scenarios will be presented for review. W19 Understanding the Psychopath: The Theoretical and Conceptual Issues Related to Psychopathy and Their Practical Application to Understanding Violent Criminals and Their Behavior Mary Ellen O’Toole, PhD*, Federal Bureau of Investigation, Behavioral Analysis Unit, FBI Academy, Quantico, VA 22135; and Robert D. Hare, PhD*, University of British Columbia, #26 15020 27A Avenue, Surry, British Columbia V4P 2Z9, Canada Upon completion of this workshop, the participant should be able to better understand the theoretical, conceptual and operational issues associated with psychopathy and its implications for criminal justice and mental health. Particular attention will be paid to the application of the theory and research on psychopathy to law enforcement, crime scene analysis, and criminal investigation and interrogation. Participants will learn about clinical and non-clinical methods for the assessment of psychopathy and its association with recidivism and violence. They also will learn how knowledge about psychopathy can be applied to crime scene analyses, criminal investigations, hostage negotiations, and interviews and interrogations. Psychopathy has emerged as one of the most important clinical constructs in the mental health and criminal justice systems. Although psychopaths make up only about 15-20% of the criminal population, they may be responsible for more than half of the serious, violent, and repetitive crimes. This presentation will impact the forensic community and/or humanity by assisting in the ability to distinguish between psychopathic and other offenders, and to understand the implications of this distinction, is critical for everyone involved in criminal justice. Dr. Robert Hare has researched psychopaths for more than a 35 years and is recognized as the world’s foremost expert on the topic. Psychopathy has emerged as one of the most important clinical constructs in the mental health and criminal justice systems. The psychopath is defined by a unique constellation of affective, interpersonal, lifestyle and antisocial characteristics. These include egocentricity, manipulativeness, callousness, impulsivity, shallow emotions, grandiosity and lack of remorse for one’s actions. Not every psychopath will have contact with the criminal justice system. However, the traits associated with this personality disorder place psychopaths at risk for committing crime and for acting out violently. Psychopathy is not limited to a remote murder scene in the middle of a desert. Psychopaths can also be found in board rooms, and government and political offices throughout the world where they wreak havoc on co-workers, investors, clients, and others with whom they come into contact. Psychopaths and their crimes are different from those of other offenders: more violent, cold-blooded, and persistent. Their lack of conscience, remorse and empathy readily allows them to become human predators, and to engage in instrumental aggression without concern for the victims or others around them. It is critical for those working in the criminal justice system to understand and appreciate the significance of this distinction between psychopathic and other offenders. An assessment of psychopathy is predictive of response to therapy, recidivism, violence, and sexual offending. The combination of psychopathy and deviant sexual arousal is particularly dangerous to society. This workshop will be two faceted: Information will be presented regarding both the theoretical and conceptual issues related to psychopathy, including current research on psychopathy as a risk factor for recidivism, violence and sexual offending. The assessment instruments for Psychopathy will be discussed including the Hare Psychopathy Checklist (PCL-R), the Screening Version (PCL-SV) and Youth Version (PCL-YV). Shooting Reconstruction, Trajectory, Bloodspatter 26 * Presenting Author In Part II, information will be presented which “operationalizes” the psychopathy construct in terms of a crime scene behavior classification system. This perspective on psychopathy will identify some key behaviors observed at a violent crime scene which can be indicative of a psychopathic offender. Utilization of this type of behavior classification system can assist law enforcement in developing strategies for investigations, interviews and prosecution of cases involving psychopathic offenders Psychopathy, Violence, Crime Scene Behavior W20 The Uses and Abuses of Statistics and Fordisc in Forensic Anthropology Richard L. Jantz, PhD*, University of Tennessee, Department of Anthropology, 252 South Stadium Hall, Knoxville, TN 37996; Stephen Ousley, PhD*, Smithsonian Institution, Director, Osteology Laboratory, Repatriation Office, Department of Anthropology, NMNH MRC 138, Washington, DC 20013-7012; and M. Kate Spradley, MA*, and Nicholas Hermann, PhD*, University of Tennessee, Department of Anthropology, Knoxville, TN 37996-0720 Upon completion of this workshop, the participant should be able to choose among several alternatives of statistical analysis based on the strengths and weaknesses of each method. The participant will have been instructed in measurement techniques and will be better able to address the reliability of a statistical analysis, both his or her own as well as others' analyses. Forensic anthropologists increasingly use FORDISC and other statistical techniques to analyze remains and make probablistic statements concerning ancestry, sex, stature, and other individuating characteristics. This presentation will impact the forensic community and/or humanity by making the attendees aware of the theory and assumptions underlying each method in order to choose the best method, to make better estimates, and to address reliability concerns. Various statistical procedures, especially discriminant function analysis (DFA), are being used more and more in Forensic Anthropology. Several computer programs can be used for statistical analysis, including SAS, SYSTAT, and FORDISC. The power of DFA lies in the fact that DFA maximizes the differences in bone size and shape among groups. FORDISC 2.0 has been used extensively to aid in ascertaining the biological profile of skeletal remains, especially for ancestry. The impetus to develop FORDISC came from increasingly frequent requests from forensic anthropologists to calculate "made-to-order" discriminant functions (DFs) using data from the Forensic Data Base (FDB). Such custom DFs are necessary when measurements required by published DFs, for example Giles and Elliot (1962; 1963) or Jantz and Moore?Jansen (1988), are impossible to obtain. DFs are also desirable when one wishes to compare the unknown to different reference groups. FORDISC allows anthropologists to construct DFs using two to eleven modern groups, some including males and females, using up to 34 craniometrics, or two to four groups using up to 39 postcranial measurements. Fordisc uses Linear Discriminant Functions (LDFs) to calculate probabilities of group membership and is easy to use. Perhaps it is too easy to use. In our experience, many users merely type in measurements and record the classification. However, there are a number of statistical assumptions and caveats that need to be addressed in DFA to avoid certain problems, the most consequential being an incorrect classification of remains. The appropriateness of certain samples should also be considered. For example, LDFs require approximate multivariate normality and about the same level of variation among groups in order to produce reliable results. If group levels of variation differ greatly, Quadratic Discriminant Functions or non-parametric DFA are solutions. Additionally, much research has been devoted to ascertaining the "best" variables to use in DFA. Noise variables do not aid in * Presenting Author discrimination and can decrease the accuracy and reliability of DFA. Stepwise selection of variables is valuable with some cautions. These methods are available in several statistical software packages and will gradually be incorporated in the next version of FORDISC. Additionally, FORDISC 2 enabled the easy estimation of stature from long bone lengths. There are also several cautions worthy of note in estimating stature. Attendees will hear lectures on theory, see empirical results that have practical implications, including case studies, receive instruction in craniometric measurement techniques, learn about emerging powerful new methods, and analyze example cases FORDISC, Statistical Analysis, Physical Anthropology W21 Preparing a Forensics Science Laboratory for Accreditation Under ISO 17025 Thomas J. Janovsky, BA*, Drug Enforcement Administration, Office of Forensic Sciences, 2401 Jefferson Davis Highway, Alexandria, VA 22301; Joseph P. Bono, MA*, Drug Enforcement Administration, Office of Forensic Sciences, 2401 Jefferson Davis Highway, Alexandria, VA 22301; Vickie A. Bailey, MS*, Northeast Laboratory, 99 Tenth Avenue, Suite 721, New York, NY 10011; Scott R. Oulton, BS*, Department of Justice, Drug Enforcement Administration, Vista, CA; Jerry A. Walker, MS*, U.S. Drug Enforcement Administration, Mid-Atlantic Laboratory, 1440 McCormick Drive, Largo, MD 20774; Donald A. Wyckoff, BA, BS*, 209 East Lewis, Pocatello, ID 83201 Upon completion of this workshop, the participant should understand how to prepare a laboratory or laboratory system for accreditation under ISO 17025 Standards. The preparation of a laboratory system is more demanding than the preparation of an individual laboratory because of the coordination required among the Quality Managers in all laboratories. Different approaches to problem solving must be harmonized to meet the requirements for the highest quality forensic work product. This presentation will impact the forensic community and/or humanity by examining the experiences of one large laboratory system in preparing for an accreditation under ISO 17025 standards, forensic laboratory managers contemplating a similar accreditation status will be exposed to the successes and potential hurdles in the process. The objective of this workshop is to present a synopsis of the procedures which were developed to prepare the nine (9) Drug Enforcement Administration (DEA) Office of Forensic Sciences laboratories for accreditation under ISO/IEC 17025 General Requirements for the competence of testing and calibration laboratories. Accreditation is becoming a de facto requirement for the recognition of a laboratory's operation under documented policies and procedures. Three states have mandated laboratory accreditation for testifying in courts. The Drug Enforcement Administration (DEA) laboratory system was first accredited in 1994, and reaccredited in 1999. These accreditations were conducted under the standards and criteria of the American Society of Crime Laboratory Directors/Laboratory Accreditation Board. In 2002 management within the DEA Office of Forensic Sciences made the decision to pursue the 2004 accreditation of the laboratory system under a program which would use as a base the International Standard ISO/IEC 17025 General Requirement for the competence of testing and calibration laboratories. Preparation for the 2004 accreditation of the DEA laboratory system would involve the Quality Assurance Managers from the seven regional laboratories, the Special Testing and Research Laboratory, and the Digital Evidence Laboratory. The accreditation also involved assessment visits to two (2) subregional laboratories, and one mobile laboratory. These three facilities are under the operational control of a regional laboratory director. The Office of Forensic Sciences Quality Assurance Manager had the 27 overall responsibility for coordinating the preparation of these twelve facilities for the accreditation process. The assessments included the following forensic disciplines: drugs, fingerprint comparisons and identifications, toolmarks examinations on solid dosage units, and digital evidence. Accreditation of a laboratory system with a significant number of laboratories spread across the United States required planning and coordination. Since the 2004 accreditation would be conducted under a set of standards different from the two previous accreditations, significant training of personnel from each laboratory was required to familiarize individual laboratory management with the requirements of ISO and the accrediting body. The following is a synopsis of the steps which brought the accreditation process to a successful completion: • The appointment of a Quality Assurance Manager within the Office of Forensic Sciences to coordinate the accreditation process. • The familiarization of Quality Assurance Managers within each laboratory with the standards for an ISO accreditation program. • Internal pre-accreditation inspections of all laboratories. • Periodic meetings of key individuals within the laboratory system to discuss and develop responses to each of the requirements for accreditation. This involved enhancing the laboratory system's policies and procedures to meet the requirements. • The compilation of a conformance file to address each individual requirement for accreditation under an ISO program. In June 2004, the Federal contract for accreditation of the DEA laboratory system was awarded to the American Society of Crime Laboratory Directors/Laboratory Accreditation Board. (ASCLD/LAB). The ASCLD/LAB International Program provides accreditation of forensic sciences laboratories under ISO/IEC 17025 requirements, enhanced by ASCLD/LAB-International Supplemental Requirements. The latter are based on the elements of the ASCLD/LAB Legacy Accreditation Program and ILAC Guide 19. After award of the contract to ASCLD/LAB, the preparation of the DEA laboratory system continued with a focus on meeting the specific requirements of the accrediting body. An important part of this workshop will involve detailed discussions of those accreditation requirements which caused a significant degree of concern and fell outside of the "comfort zone" of familiarity with previous accreditation inspections. The discussions in this workshop will be candid. The goal of the organizers is to present suggestions for developing viable, practical suggestions for achieving accreditation under an ISO accreditation program for a forensic science laboratory. Laboratory Accreditation, ISO/IEC 17025, Drug Enforcement Administration W22 Evidence-Based Forensic Science: Interpreting Postmortem Toxicology in the Light of Pathologic Findings Paul Fornes, MD, PhD*, Anatomie Pathologique, Department of Pathology, Hopital Europeen Georges Pompidou, 20, rue Leblanc, Paris, 75015, France; Barry K. Logan, PhD*, Washington State Patrol, 2203 Airport Way, SOuth, #360, Seattle, WA 98134-2027; Marc D. Bollmann, MD*, Institut Universitaire de Médecine Légale, Rue du Bugnon 21, Lausanne, CH-1005, Switzerland; Olaf Drummer, PhD*, Monash University, Department of Forensic Medicine, Victoria, 3800, Australia; A. Robert W. Forrest, MB ChB, LLM*, University of Sheffield, Department of Forensic Pathology, Medico-Legal Center, Watery Street, Sheffield, South York S3 7E5, United Kingdom; Bruce A. Goldberger, PhD*, University of Florida, College of Medicine, 4800 SW 28 35th Drive, Gainesville, FL 32608; Graham R. Jones, PhD*, Office of the Chief Medical Examiner, 7007-116 Street, NW, Edmonton, Alberta T6H 5R8, Canada; Steven B. Karch, MD*, PO Box 5139, Berkeley, CA 947050139; Christopher M. Milroy, MD, PhD*, University of Sheffield, Department of Forensic Pathology, Medico-Legal Center, Watery Street, Sheffield, South York S3 7E5, United Kingdom; Ashraf Mozayani, PharmD, PhD*, Harris County Medical Examiner's Office, 1885 Old Spanish Trail, Houston, TX 77054; and Derrick J. Pounder, MD, ChB*, University of Dundee, Scotland, Department of Forensic Medicine, Dundee, Scotland DD1 4HN, United Kingdom After attending this presentation, this joint Toxicology/ Pathology/Biology workshop will provide both toxicologists and pathologists with guidelines in interpreting toxicology in the light of pathological findings in assessing the cause of death. Evidence-based medicine has become one of the major driving forces in clinical practice, with an impact on education, policy making and research. Evidence-based medicine is the conscientious, explicit, and judicious use of current best evidence in making decisions about the care of individual patients. The practice of evidence-based medicine means integrating individual clinical expertise with the best available external clinical evidence from systematic research (1). Can this new paradigm be applied to forensic sciences, in particular to forensic toxicology and pathology ? As far as the determination of the cause and manner of a death is concerned, the presence of a substance in a body raises the question of its possible role in the death. Attempts at directly relating postmortem blood drug concentrations to outcomes only seem reasonable to those unaware of just how many variables need to be considered. In addition to a thorough account of what was observed at the scene and an inquiry into the decedents past medial history, the medical examiner should integrate both toxicological and pathological findings. It is the purpose of this joint toxicology/pathology workshop to provide the forensic physician with the most advanced and objective knowledge in both toxicology and pathology, in accordance with principles of forensic-based medicine. The following presentations will examine toxicological and pathological issues of practical relevance to the assessment of the cause of death. 1. Introduction. The principles of evidenced-based medicine. 2. Sampling guidelines for toxicology. 3. Hair Testing : Why Postmortem Toxicology is Incomplete Without it. 4. Back Calculation, is the Process Ever Legitimate in the Dead ? 5. Postmortem Redistribution 6. Influence of pharmacogenetics and impaired metabolism in chronic accumulation of medications 7. Interpreting Toxicology in the Light of Pathologic Findings a) Cardiac Pathology b) Other Pathologic Findings 8. Alternative Explanations for Sudden Cardiac Death, an Introduction to the Molecular Biology of Ion Channel Disease. 9. Interpreting Postmortem Opiate Measurements ; Methadone, Oxycodone, Bupranorphine, and Other Substitutes Drugs. 10. Interpreting Postmortem Stimulant Measurements: Cocaine, Methamphetamine, and MDMA 11. Determining the Role of Doping in Sport-Related Deaths. 12. Special Situations a) Murder by Succinylcholine b) Murder by Epinephrine c) Euthanasia - Does Toxicology Testing Help? d) Is Murder by Breast Feeding Possible ? 13. Illustrative(s) Case(s) 14. Discussion (1) Sackett DL, Rosenberg WMC, Gray JAM, Haynes RB, Richardson WS. Evidence-based medicine: what it is and what it isn't. BMJ 1996; 312: 71-2. Toxicology, Pathology, Cause of Death * Presenting Author W23 Quality Assurance in Forensic Anthropology Vincent J. Sava, MA*, Andrew J. Tyrrell, PhD*, and Thomas D. Holland, PhD*, JPAC CIL, 310 Worchester Avenue, Building 45, Hickam AFB, HI 96853 After attending this presentation, attendees will understand the basic quality assurance principles and measures applicable to forensic anthropologists. Participants will learn the unique challenges faced by forensic anthropologists when striving to have their facilities, procedures, and casework meet the standards demanded by the criminal justice system. Attendees should be able to utilize the material presented to formulate a quality assurance program for their organization. Quality assurance in forensic laboratories and programs has become a growing trend over the past decade. This presentation will impact the forensic community and/or humanity by demonstrating how formal quality assurance programs can lead to objective and measurable standards and performance that ultimately strengthen and elevate the forensic science profession as a whole. Quality assurance programs in forensic laboratories and activities have been a growing trend over the past decade. Since 1999 the Joint POW/MIA Accounting Command, Central Identification Laboratory (JPAC-CIL) has implemented a stringent quality assurance program to ensure the scientific integrity of its casework. The CIL's quality assurance program ultimately led to the Laboratory's accreditation by the American Society of Crime Laboratory Directors Laboratory Accreditation Board (ASCLD-LAB) in 2003--the first forensic skeletal identification laboratory to be so credentialed. The goal of this workshop is to introduce the attendee to the CIL's Quality Assurance Program and to convey the lessons learned resulting from its implementation and growth. A video overview of the JPAC CIL is presented followed by an overview of its quality assurance program. In the latter, the concept of the scientific integrity of the laboratory is discussed followed by a summary of the "Surety" model of quality assurance. The participants will become familiar with each measure that comprises the surety model of quality assurance. The importance of integrating and synchronizing all of the surety measures discussed during the workshop will be continually stressed. In Part I, infrastructure and support considerations necessary for a successful quality assurance program are also presented. Surety measures addressed include: • Desired qualities of a laboratory manual and other vital documentation • Adequacy and safety of facilities • Policies and procedures conducive to a positive work environment • Evidence management and security • Training and professional development Gathering and interpreting evidence is the focus of Part II where quality assurance in field operations and trace evidence analysis is discussed. The surety measures directly related to casework--the peer review process, validation of technical procedures, case file management, analytical notes and documentation--are presented for consideration. Quality assurance procedures and programs are ineffective in the absence of monitoring, enforcement, and corrective action. These are accomplished through a myriad of surety measures including proficiency testing, review of court testimony, audits, annual reports, and corrective action policies, which are presented in Part III. In closing, the attendees will become acquainted with the problems that hindered, and the processes that led to, the accreditation of the JPAC CIL. In closing, surety assistance programs offered by the CIL will be discussed in the event an attendee's organization desires assistance with their surety programs or accreditation efforts. A question and discussion period follows the conclusion of the presentation portion of the workshop. Quality Assurance, Forensic Anthropology, Accreditation * Presenting Author W24 The Forensic Nurse Death Investigator as a Member of the Multidisciplinary Forensic Investigative Unit Donna M. Garbacz Bader, MSN, RN*, Bryan LGH, College of Health Sciences, 5035 Everett Street, Lincoln, NE 68506; Virginia A. Lynch, MSN, RN*, Beth EL College of Nursing, University of Colorado, 514 Hopi Circle, Divide, CO 80814; Vincent J.M. DiMaio, MD*, Bexar County Medical Forensic Science Center, 7337 Louis Pasteur, San Antonio, TX 78229-4565; and Stacey Mitchell, MSN, RN*, and Luis Arturo Sanchez, MD*, Office of the Medical Examiner of Harris County, 1885 Old Spanish Trail, Houston, TX 77054 Upon completion of this workshop the participant will be able to identify and discuss the correlation of scientific nursing education to the field of forensic science, specifically to that of death investigation. This presentation will impact the forensic community and/or humanity by increasing the availability of knowledgeable, competent individuals to assist the healthcare and legal system in relevant assessment and documentation in the field of death investigation. Death investigation is a complex process, one that involves accurate data collection, communication and documentation. As a science dealing in an objective assessment of death, the investigative process requires a degree of knowledge in human anatomy and physiology, psychology, basic chemistry and physics. A variety of disciplines are essential in order to accurately assess, interpret and correlate the various elements involved in decedent identification, cause and manner of death, support and encouragement to survivors, and to maintain channels of communication with additional investigative personnel. Members of the multidisciplinary death investigative team have routinely included law enforcement agencies, crime laboratory personnel, and medical examiner and/or coroner (ME/C) investigators. Additional team members, include those specializing in forensic engineering, odontology, anthropology and the behavioral sciences. Forensic Nurse Death Investigators (FNDI) have recently become valued members of death investigative systems and are increasing in many ME/C jurisdictions. The educational requisites of the FNDI include specific aspects of each of these various forensic disciplines with which they regularly interface. Forensic nurse examiners (FNE) in the United States are recognized for their clinical education and experience in the biomedical sciences. The FNE is accomplished in the application of scientific process to objective data collection, evaluation and interpretation of that data, including the implementation of methods essential to the investigation of suspicious deaths. Forensic nursing science requisite skills involve observation, documentation and assessment of objective data, human anatomy and physiology, basic chemistry and biochemistry, microbiology, physics and pharmacology. Familiarity with human psychology in response to both internal and external stressors helps to prepare the FNE for the task of death notification. In addition, an understanding of cultural family systems and available community resources is essential in order to provide direction and support to the bereaved. Thus the FNE is uniquely qualified to fill the role of the forensic death investigator. ME/Cs who employ FNDIs appreciate the educational requirements for an individual to become a licensed professional. Scientific based education and critical thinking skills allow the forensic nurse examiner to recognize and recover physical and biological evidence with laboratory specification. The ability to identify specific human elements basic to normal physiological and psychological functions across the life span as well as assist the forensic nurse examiner in the identification of trauma in human remains. Considering that the majority of all reported ME/C cases are comprised of natural deaths, the biomedical expertise of the FNDI provides for differentiation between disease process and suspicious deaths with 29 reasonable certainty. One emerging application of the FNE in death investigation is providing postmortem sexual assault examination prior to autopsy. Forensic pathologist and the FBI criminal laboratory find the FNE exceptionally qualified as experts in the evaluation of sexual assault trauma and collection of biological evidence. A question frequently asked is: What qualifies the Registered Nurse to participate in death scene investigations, postmortem procedures and provide expert witness testimony? To examine the evolution of a forensic specialist in nursing, one must first consider the initial education, which begins as a nurse matriculates through an accredited nursing program. Basic nursing curricula focus on a strong physical and psychological science base, including human anatomy and physiology, chemistry, biology, basic and advanced mathematics, physics, behavioral sciences, and general courses in the fine arts. The scientific process is applied throughout all aspects of general nursing curricula; the basic sciences are correlated to various body systems and related natural disease. Legal and ethical issues are a major component of each course. Graduates are eligible to apply for licensure examination developed by the National Council of State Boards of Nursing. The successful candidate is awarded the designation of Registered Nurse and licensed to practice, independent of physician licensure. At this juncture, nurses with an interest in the forensic sciences seek out accredited forensic nursing science programs. In recent years, undergraduate, graduate, and postgraduate programs in forensic nursing science have been developed and are continuing to develop across the U.S. and abroad. These programs offer specific forensic curricula in the scientific investigation of injury and death, human abuse, forensic chemistry, crime scene/crime laboratory, forensic photography, toxicology, victimology, traumatology, sexual violence, human rights, psychosocial and legal aspects of forensic science, among others. Advanced forensic nursing curricula pursues pathophysiology, research, epidemiology, consulting, curriculum design and informatics. As generally happens, the greater desire for knowledge promotes higher academic goals. To date, four accredited United States (U.S.) universities offer doctorate level forensic nursing degrees. As principle research investigators and associates to forensic pathologists, it is believed that future advance practice forensic nurse examiners will assist in alleviating a global shortage in competent forensic services pertaining to the scientific investigation of death. In tribute to the AAFS, forensic nursing was first recognized as a scientific discipline at the 43`d annual meeting in 1991 (Anaheim, California). The International Association of Forensic Nurses (IAFN), patterned after the AAFS, was founded in 1992. In 1995 the American Nurses Association (ANA) Congress of Nursing Practice bestowed formal recognition to forensic nursing as an official nursing specialty, and in 1997, the IAFN published the Standards and Scope of Forensic Nursing Practice in conjunction with the ANA. IAFN board certification for the forensic nurse examiner in both adult and pediatric sexual assault examination and evaluation was implemented in 2001. Development of specific standards and national certification in biomedical investigation for the Forensic Nurse Death Investigator is currently in progress through the IAFN. This certification will include professional standards in postmortem sexual assault examination for those who practice in this field. The utilization of the FNDI provides competent individuals skilled in the biomedical investigation of death. Where perpetrator prosecution may be indicated, successful resolution to questioned death related issues instills confidence in the health and justice systems and contributes to community mental health through social justice. This workshop will address the development of forensic nursing education programs, certification examination, role clarification, investigative case management, and multidisciplinary team relationships. Case examples are presented in which the FNDI's biomedical skills benefit the forensic pathologist in the analysis of fatal injuries and medical deaths. The foremost FNDI investigative program in the U.S. will present an overview of the Medical Examiners Investigative Unit comprised exclusively of Forensic Nurse Death Investigators. Discussion of the 30 unlimited potential for FNDIs in developed and developing countries is previewed with data related to existing programs and those currently developing. Death Investigation, Forensic Nurse Death Investigator, Forensic Nurse Examiner (FNE) W25 Identifying Printmaking Techniques Through the Artistic Process Troy J. Eberhardt, BS*, Department of Homeland Security, Immigration & Customs Enforcement, 800 Westpark Drive, Suite 325, McLean, VA 22102-3105; William J. Farrell, Jr., MS*, Forensic Consultant, PO Box 6974, Metaire, LA 70009; Anna C. Baskin, MA*, 7619 Burthe Street, New Orleans, LA 70118; and Peter Belcastro, MS*, and Gabriel Watts, BS*, FBI Laboratory, 2501 Investigation Parkway, FBI Academy Complex, Quantico, VA 22135 The goal of this workshop is to provide for a better understanding of scientific techniques, applications and future implications on the printmaking industry. Ideally, the scientific community will gain a more accurate and detailed understanding of the processes used in printmaking so that detection and analysis will be better accomplished. This presentation will impact the forensic community and/or humanity by providing the forensic document examiner a greater understanding of printmaking through fine arts processes and gain knowledge of the cross over between industrial printing and fine art printing. The attendees of this workshop will be introduced to the artist techniques and the ongoing use/development of Printmaking. Printing techniques, inks and paper are continuously being updated over time by artists as well as the printing industry. A discussion of the history of printmaking will be presented along with ideas of future techniques in the printing industry. This program will consider the traditional methods of printing (lithography, intaglio, wood cut relief, silk screen, pad printing, photo polymer plates, letter press) as well as future implications of computer applications, digital, and holography printing. Printmaking, Artist Techniques, Traditional Printing WS1 Filicide: Risk Factors and Psychological Aspects Karen F. Ross, MD*, Jefferson Parish Forensic Center, 2018 8th Street, Harvey, LA 70058; and John W. Thompson, Jr., MD*, Tulane University School of Medicine, Department of Psychology and Neurology, TB53 1440 Canal Street, 10th Floor, New Orleans, LA 70112 The goal of this presentation is to define filicide and related forms of child abuse; list the 3 general categories of fatal child abuse; list previous classifications of filicide; list maternal, paternal and infant risk factors for filicide; discuss psychological aspects of filicide including motivating factors in the absence of insanity; note the differences between filicide perpetrators and other perpetrators of child abuse. This presentation will impact the forensic community and/or humanity by discussing filicide in a way that will strive to enlighten those that investigate these tragic cases about the risk factors in the parents and the child, and motivating factors in the absence of insanity as well as approach to evaluation of psychological make-up of the perpetrator in hope of providing preventive measures and improve the adjudication of such cases. * Presenting Author Filicide is the killing of a child by a parent at any age. While all child abuse cases are tragic, none provoke more questions and emotional response than the case of a child (or sometimes multiple children) who is killed by a parent. This may result at least in part because of the incongruity between societal expectations that parents should love, nurture and protect their offspring and the reality in such cases that they brutally abuse and murder them. This workshort begins with an overview of the broad categories of child abuse (gentle homicide, battered child Syndrome, and Impulse Homicide), then offers a review of previous classifications of filicide, discusses risk factors for filicide including motivating factors and psychological aspects and addresses ways in which filicide perpetrators differ from other perpetrators of child abuse including increased likelihood of mental illness. Differences in motivating factors and psychological features of the perpetrators will be discussed in context of the 3 broad categories as listed above also. Filicide, Child Abuse, Psychological Evaluation poisoning explained the course of Napoleon's entire illness, with the exception of the last several weeks, when he believes the emperor also suffered from antimony poisoning as a result of too much tartar emetic and from mercury poisoning as a result of too much calomel. Forshufvud explained his hypothesis in a 1961 book titled Who Killed Napoleon? He later obtained one of the hair strands that had been cut from Napoleon's head the day after his death. This piece of hair was analyzed for arsenic by neutron activation analysis, at the British Atomic Energy Establishment in Harwell. The results, which were published in 1962 in the journal Nature, showed that the hair contained more than 10 parts per million of arsenic, well over 10 times the normal value. Other specimens of Napoleon's hair have also been proven to contain arsenic. However, the cause of Napoleon's death is still being debated by historians. Finally, the following issues will be discussed: Was arsenic a contaminant, or was it inhaled or ingested, and was it actually the cause of death? If Napoleon did have gastric carcinoma, did he die of it or with it? Was Napoleon murdered? By whom? How would have the murderer used arsenic? What could have been the motive? Napoleon, Hair Testing, Arsenic WS2 Latest Advances in Hair Testing and Cardiac Electrophysiology Applied to the Determination of the Cause of Death of Napoleon Bonaparte Paul Fornes, MD, PhD*, Department of Pathology, Hospital E.G. Pompidou, 20, rue LeBlanc, Paris, 75015, France; and Steven B. Karch, MD*, PO Box 5139, Berkeley, CA 94705-0139 After attending this presentation, attendees will be able to examine comparatively the results obtained by different technologies : Instrumental Neutron Activation Analysis, Graphite Furnace Atomic Absorption Spectroscopy, Inductively Coupled Plasma Mass Spectrometry, Electrothermal Atomic Absorption Spectrometry, Synchrotron and X-Ray fluorescence, and Nano-Secondary Ion Mass Spectrography. Analytical procedures and decontamination methods will be examined. Limitations of these methods will be discussed; and will be able to examine the possible roles of arsenic, mercury chloride (calomel), and tartar emetic (antimony potassium tartrate), in the light of most recent cardiac electrophysiological studies. These findings will be discussed in the light of autopsy findings and historical background. The present forensic works will impact the forensic community and/or humanity by contributing to improve our knowledge of history. On May 5, 1821, Napoleon Bonaparte died at the age of 51, on Saint Helena island, where he was detained in exile since October 1815. An autopsy was performed by Francesco Antommarchi, Napoleon's personal physician, witnessed by seven British medical officers. Antommarchi's report described among other findings, a perforated gastric ulcer contained by adhesions to the liver. The gastric lesions were considered cancerous, but there was no peritonitis, nor peritoneal carcinosis, nor visceral metastases. Yet, most history books state that Napoleon died of cancer of the stomach. In accordance with his wishes, Napoleon's hair was then cut off and strands were distributed as keepsakes to the various members of his establishment. In 1840, Napoleon was exhumed on the order of the French government, and the remains were then brought to Paris, where they now rest in the Hotel des Invalides. After studying various reports, a Swedish dentist named Sten Forshufvud concluded in the early 60s that acute and chronic arsenic * Presenting Author WS3 Chemistry of Voodoo: Murder in the French Quarter Richard C. Froede, MD*, 3930 North Placita de la Escarpa, Tucson, AZ 85750; Frank Minyard, MD*, New Orleans Coroner Office, 2700 Tulane Avenue, New Orleans, LA 70119; Walter H. Birkby, PhD*, Forensic Science Center, 2825 East District Street, Tucson, AZ 85714; Homer R. Campbell, Jr., DDS*, Office of the Medical Investigator, MSC11 6030, 1 University of New Mexico, Albuquerque, NM 871310001; and Suzanne M. Froede, MA*, 3930 North Placita de la Escarpa, Tucson, AZ 85750 Upon completion of this presentation, attendees will understand the fundamentals of crime scene investigation; understand the need for team cooperation in death investigation; appreciate the procedures followed in a Coroner’s court; and understand the practice of “voodoo” and it’s chemistry. Welcome to New Orleans, a fabulous city for fun, frolic and mystery. It’s a city with wonderful restaurants and tantalizing food; great jazz and Cajun music; moonlight nights on the Mississippi; carriage rides; marvelous coffee and beignets; Bourbon Street; hurricanes; and Voodoo. New Orleans also has a seamy/steamy side; muggings, scams; corruption; and MURDER! An opportunity will be afforded to the amateur detective to analyze and solve a series of murders. The participants to analyze and collect evidence will use on-scene investigative techniques. The participants; powers of observation and knowledge of pathology will be challenged to determine the cause of death and to find the killer. A police detective and medical examiner will be there to assist you, but you must be careful of the representatives of the media. The objectives in these cases are to develop skills in analyzing and reconstructing a crime(s) as well as to determine motivation in cases where there is conspiracy and complicity. Investigation, Chemistry and Toxicological, Coroner’s Court Procedures 31 Criminalistics B1 Evaluation of Y-STR Multiplexes for Analysis of Casework Samples Lora Bailey-Van Houten, BS*, Office of the Attorney General, Bureau of Forensic Services, Fresno Regional Laboratory, 5311 North Woodrow Avenue, Fresno, CA 93740; and Sulekha R. Coticone, PhD, California State University, Department of Chemistry, 2555 East San Ramon Avenue, Fresno, CA 93740 After attending this presentation, attendees will learn the relative abilities of various Y-STR multiplexes for forensic use. This presentation will impact the forensic community and/or humanity by assisting in the ability to determine relative performance of Y-STR multiplexes. Fluorescence-based DNA detection systems are being widely in forensic DNA analysis. These methods have greatly assisted the sensitivity and ease of measurement of PCR amplified short tandem repeat (STR) alleles. In multiplexed STR genotyping kits, fluorescent dyes are covalently coupled to one primer for each locus. These STR multiplexes amplify 13 autosomal STRs with a power of discrimination of over one in a billion and have proven invaluable in identification of perpetrators of violent crimes. However, sexual assault cases often contain a mixture of DNA from the male perpetrator and the female victim that are difficult to interpret using autosomal STR kits, due to the presence of excess female DNA in these samples as compared to the male DNA. By using male polymorphisms on the Y chromosome, male DNA can be identified specifically in male-female mixed samples. The present study was conducted to evaluate two recently introduced commercial Y-STR multiplexes (Promega’s PowerPlex® Y and Reliagene’s YPlex 12). The robustness of the multiplexes was determined by sensitivity studies. The sensitivity data indicated that both kits were sensitive, although the YPlex 12 displayed consistently higher peak heights. Additional stutterlike peaks were noticed in pristine male samples at DYS392, DYS389II, DYS437 and DYS385 loci, though no artifacts were noticed when amplification was performed with female DNA. Further analysis of these artifacts need to be performed to determine the origin of these additional peaks. Mixture studies indicated that the limit of detection of the minor component in a male:male mixture was 1:5 (for YPlex 12) and 1:2 (for PowerPlex® Y). Researchers also performed CEPH family studies to demonstrate Mendelian inheritance of the Y-STR loci. To assess the ability of the multiplexes to analyze forensic samples, testing on blood, oral swabs and male-female mixtures as well as previously adjudicated sexual assault samples were performed. Based on these studies, the relative ability of the two multiplexes to successfully analyze a variety of forensic was determined. Y-STRs, Multiplexes, Casework B2 Optimization of the Extraction of Total Ribonucleic Acid (RNA) From Semen Rachel A. Bartholomew, PhD*, FBI Laboratory, FBI Academy, Counterterrorism and Forensic Science Research Unit, Building 12, Quantico, VA 22135; Jack Ballantyne, PhD, and Jane Juusola, BS, University of Central Florida, PO Box 162366, Orlando, FL 32816; Richard Guerrieri, MS, and Rhonda Craig, MS, FBI Laboratory, DNA Analysis Unit 1, 2501 Investigation Parkway, Quantico, VA 22135; and Kevin W.P. Miller, PhD, FBI Laboratory, FBI Academy, Counterterrorism and Forensic Science Research Unit, Building 12, Quantico, VA 22135 This presentation will impact the forensic community and/or humanity by demonstrating how messenger RNA expression patterns can provide cell and tissue specific information that can be used to positively identify a tissue or body fluid source. Subsequent amplification of mRNA via RT-PCR using tissue specific genes can be to identify the tissue/body of origin. Although DNA technology can be used to identify (i.e., type) a suspect, the source of DNA itself (blood, semen, etc.) cannot always be definitively identified. Therefore there is a need for the development of tissue or body-fluid specific protocols to identify forensically relevant body fluid or tissue samples. The development and use of RNA technology may complement existing DNA technology where the source of the stain (i.e., body fluid or tissue type) can be identified by examining the expression profile of body fluid or tissue-specific genes. Each cell type has a distinctive pattern of messenger RNA (mRNA) expression. Messenger RNA expression patterns provide cell and tissue specific information that can be used to positively identify a tissue or body fluid source. Subsequent amplification of mRNA via RT-PCR using tissue specific genes can be to identify the tissue/body of origin. The RNA extraction protocol was streamlined for ease of use in the forensic laboratory. The traditional RNA extraction protocol is lengthy, produces toxic bi-products (i.e., phenol), and requires a skilled user. To be validated and used in casework, stringent quality control measures must be performed on each batch of new reagents, decreasing sample throughput. The use of a RNA extraction kit has several benefits: (i) increased sample throughput, (ii) lack of toxic/hazardous chemical bi-products, (iii) ease of training, and (iv) “built in” quality control measures. The extraction of RNA from liquid body fluids was optimized using cryopreserved human semen. RNA was extracted from 0.5-500ul of semen using the RNAeasy Micro kit (Qiagen, Valencia, CA). The Agilent 2100 Bioanalyzer was used to determine RNA quantity and quality, including RNA Integrity Number (RIN) analysis. RNA samples (0.5pg to 50ng) were reversed transcribed using the Sensiscript Reverse Transcription kit (Qiagen, Valencia, CA). PCR products were amplified with semen-specific Protamine-1 primers. The subsequent PCR products were analyzed using the Agilent 2100 Bioanalyzer. The amount of total RNA extracted from semen samples ranged from 9pg/ul to 20ng/ul. RNA quality was variable, with RINs ranging from 1-8.6 (scale 1-10). RNA samples having a RIN of 1 lacked 18S and 28S ribosomal peaks, but still contained quantifiable RNA. Samples with a RIN of 8.6 contained intact 18S and 28S ribosomal RNA bands. RNA samples (degraded and non-degraded) were efficiently reverse transcribed and amplified to produce a semen specific RNA product. A semen-specific Protamine-1 RT-PCR product was obtained from as little as 5pg (RIN=7.4) or 15-25 pg (RIN=1). The amplification of a semen specific product was possible with degraded RNA, suggesting that RNA can be used for cellular source attribution even when there is a question regarding the overall integrity of the sample. The potential benefits of an RNA based approach for body fluid stain or tissue characterization include: (i) the ability to perform parallel tests for numerous markers of a single body fluid in a single assay format, (ii) the ability to perform parallel tests for different body fluids in a single assay format, (iii) a definitive identification of body fluids for which presently no specific test exists, and (iv) the ability to automate the process. The use of RNA technology could supersede current protocols for forensic body fluid identification. Serology, Semen, RNA Analysis The goal of this presentation is to optimize the extraction of total ribonucleic acid (RNA) from body fluids for reverse transcription polymerase chain reaction (RT-PCR) and subsequent determination of source attribution. 32 * Presenting Author B3 Verification of STR Alleles by Alternative Primer Pairs Through a Singleplex PCR System Eng Seng Simon Lim, BS*, Hui Koon Joyce Lai, BS, Chien Tien Grace Law, BS, Hui Koon Joyce Low, BS, and Wai Fun Tan-Siew, MS, Health Sciences Authority, DNA Database Laboratory, 7 Maxwell Road, MND Building Annex B, #05-04, Singapore, 069111, Singapore The Singapore National DNA Database Laboratory uses the ABI IdentifilerTM kit for DNA profiling of the 13 CODIS loci. Routinely, researchers encounter allele ambiguities such as off-ladder range alleles, microvariant alleles, allele peak imbalances and tri-alleles among the different DNA loci typed. In order to determine whether these are true alleles or PCR artifacts, alternative primers of the questioned DNA locus are used to confirm the alleles typed. An allele is automatically assigned by its DNA locus specific Genotyper™ macro. Using the FGA locus as an example, the singleplex PCR of the questioned DNA locus approach is described here. This presentation will impact the forensic community and/or humanity by demonstrating how the systematic singleplex PCR approach with automated allele assignment method will allow routine confirmation and resolution of STR allele ambiguities encountered during routine DNA profiling in the laboratory. PCR artifacts such as stutter products and non-template nucleotide additions and other factors such as microvariants, allele dropout, allele imbalance and mutations can arise that may interfere with the clear interpretation and genotyping of the alleles present in the DNA template. In the National DNA Database Laboratory, the Identifiler™ DNA typing kit is used in routine DNA profiling. The DNA is extracted and amplified from blood stained FTA™ collection cards of convicted offenders. Duplicate DNA typing is performed for each sample and results are compared for consistency and quality before uploading into the CODIS system. On some occasions, heterozygote allele peak imbalances, three banded allele patterns, off-ladder range alleles, microvariant alleles and spurious PCR artifacts are encountered. In order to resolve interpretation difficulties and allow genotyping of the correct STR alleles present in the DNA template, alternative fluorescent (6-FAM) labelled primer pairs using Promega® Powerplex™ 16 or mini STR primer (Butler et al. 2003) sequences of each DNA locus are obtained. For each DNA locus, amplifying the alleles obtained from either the diluted Identifiler™ or Powerplex™ 16 allelic ladders as template creates the allelic ladder. A Genotyper™ macro is then written for automated allele assignment based on the amplified ladders for each DNA locus. The questioned DNA samples are then amplified using the DNA locus-specific singleplex PCR primers. Alleles are assigned on the PCR products using its DNA locusspecific Genotyper™ macro. The identity of the PCR products is verified as either as PCR artifacts or as true allele amplified from the DNA templates. An example of this approach using the FGA locus is discussed here. The Powerplex™ 16 primers for the FGA locus are used. Out of the 70 blood samples with allele peak ambiguities observed using the Identifiler™ DNA typing kit, 29 samples were confirmed to be off-ladder range alleles, 30 samples with peak imbalances were corrected and 10 tri-alleles were verified. The corrected imbalanced peak indicated point mutations at the primer binding position of the Identifiler™ FGA primers. Off-ladder range alleles were confirmed to be alleles of the FGA locus. The remaining sample with an imbalance peak typed using the Identifiler™ DNA typing kit is likely to be a tri-allele pattern as the imbalance peak was consistent with the singleplex PCR result. The tri-allele should consist of a homozygote allele peak with an imbalanced third allele. In conclusion, this singleplex PCR approach with automated allele assignment allows routine confirmation and resolution of STR allele ambiguities encountered during routine DNA profiling in the laboratory. B4 The Effects of Different Environmental Factors on Quantity of DNA Extracted From Skeletal Remains Recovered From Gravesites in Former Yugoslavia and Quality of DNA Typing Results Arijana Selmanovic, BS*, Ana Milos, BS, Jon M. Davoren, MS, Tony Donlon, BS, Lejla Smajlovic, BS, Edina Omerovic, Rijad Konjhodzic, BS, and Jon Sterenberg, MS, International Commission on Missing Persons, Alipasina 45a, Sarajevo, 71000, Bosnia and Herzegovina The goal of this presentation is to present data which will reflect upon the quality of the obtained STR profiles and the quantity of DNA extracted from bone samples taken from skeletal remains that were exposed different environmental influences, and the observed differences between them. This presentation will impact the forensic community and/or humanity by providing information to persons performing DNA STR testing of skeletal remains from mass graves. The International Commission on Missing Persons has been tasked with the challenge of identifying mortal remains from the armed conflicts in the former Yugoslavia that occurred in the 1990s. This process is complicated due to several factors: at least nine years have passed since the conflicts ended, there are up to 30,000 missing persons in graves scattered throughout the former Yugoslavia, the conditions of the mortal remains being recovered, and because DNA testing has become the only reliable means of identification in the majority of these cases. In the ICMP’s identification efforts, bodies have been found buried in mass graves of up to hundreds of bodies. Within the same grave remains are often exposed to a number of different conditions as some were buried in body bags, others were wrapped in plastic and some were in direct contact with either soil or with other bodies. Many sets of mortal remains have also been burned prior to being placed in the mass graves. DNA testing of skeletal remains is a rather challenging task because the DNA in such bone samples is generally highly degraded. In addition, it is normal that a substantial microbial population infests the bone samples. The process is further complicated because of the diverse storage conditions in which bodies were placed at time of death. The ICMP has developed a DNA-led identification process that is successful in obtaining STR profiles in over 85% of the skeletal cases from the former Yugoslavia. In order to further optimize this testing process the authors are investigating the effects of the environmental conditions of the mass graves to see if there is any link between those conditions and the recovery of DNA from bones. To determine the effects of environmental factors on the degradation of DNA, multiple bone samples from two different disposal sites were examined. The pH values of soil samples from those two gravesites were measured. Survey data of gravesites was collected with information on depth and position of bodies within the grave. Factors pertaining to conditions of bodies such as: whether bodies were burnt, whether plastic sheeting was used to wrap bodies and whether bodies were buried in body bags were also noted and considered. The relative DNA content of bone samples from those specific locations was determined by quantification of DNA extracted using The Quantifiler™ Human DNA Quantification Kit and ABI Prism® 7000 Sequence Detection System. STR Typing was performed using the commercially available Promega PowerPlex® 16 System. The data presented will reflect upon the quality of the obtained STR profiles and the quantity of DNA extracted from bone samples taken from skeletal remains that were exposed different environmental influences, and the observed differences between them. DNA, STR, Degradation STR, Peak Imbalance, PCR * Presenting Author 33 B5 Developing a Simple Method to Process Compromised Bone Fragment for Forensic DNA Isolation Richard Li, PhD*, Sandra Chapman, MS, and Mary Thompson, BS, Sam Houston State University, Box 2296, Huntsville, TX 77341; and Michal Schwartz, BS, Sam Houston State University, Box 2296, Huntsville, TX 77341 The goal of this presentation is to introduce a new method for processing compromised bone fragments prior to DNA isolation. The advantage of this method over conventional methods is in applying the proteinase solution that will omit the step of a physical cleaning procedure, such as sanding. This presentation will impact the forensic community and/or humanity by demonstrating data which suggests that this method could be used for 1) initial sample preparation for cleaning the outer surface of compromised human skeletal fragments, and 2) could be adapted for automated DNA isolation for bone fragment in the near future. Skeletal remains have been challenging biological samples for DNA isolation. Bones are more difficult for preparing and sampling prior to DNA extraction. One of the labor-intensive and time-consuming steps in DNA isolation from bone fragments is the initial cleaning and sampling of the bone. Due to the potential of having co-mingled remains, adhering inhibitors and bacterial contamination, the outer surface of the bone fragment has to be cleaned by a current method like sanding. However, to avoid cross-contamination between samples, the bone dust that is generated during the sanding of the bone must be removed. Additionally, safety protection equipment and procedures are necessary to protect lab workers and technicians from exposure to blood-borne pathogens. To address this issue, a simple processing method has been developed using proteinase solution prior to DNA isolation. In this study, the use of proteinase solution requires much less labor than a physical method such as sanding. By incubating with the proteainse solution, the soft tissue and outer surface of the bone fragment sample is removed. The processed bone fragment or a portion of the fragment can then be used for DNA isolation. The characterization of the effect of the proteinases on the cleaning of bone fragments was performed. Proteinases were screened and the best candidates were selected. Additionally, the optimum incubation condition (concentration of proteinase, incubation temperature and pH) for the proteinase was determined. DNA from processed bone sample was isolated. The results demonstrated that this method is effective for removing those soft tissues attached to bone samples and the outer surface of bone fragment samples. The data suggest that this method could be used for 1) initial sample preparation for cleaning the outer surface of compromised human skeletal fragments, and 2) could be adapted for automated DNA isolation for bone fragment in the near future. Bone, Challenging Samples, DNA Isolation B6 Investigation of the Performance of the Promega PowerPlex® 16 System for Testing of Low Copy Number (LCN) STR DNA Samples Lejla Smajlovic, BS, Edina Omerovic, Ana Milos, BS, Arijana Selmanovic, BS, Jon M. Davoren, MS, Tony Donlon, BS, and Rijad Konjhodzic, BS, International Commission on Missing Persons, Alipasina 45a, Sarajevo, 71000, Bosnia and Herzegovina After attending this presentation, attendees will be shown a number of DNA – STR profiles from the Promega PowerPlex® 16 system during validation of the system for Low Copy Number analysis. 34 This presentation will impact the forensic community and/or humanity by discussing some of the findings associated with low copy number analysis with the Promega PowerPlex® 16 system. The International Commission on Missing Persons has setup a DNAled identification system in the former Yugoslavia. The initial phase of the identification effort utilized short tandem repeat (STR) analysis to match bone samples with family reference samples. The Promega PowerPlex 16 System (PP16) has been highly successful for testing of bone samples even 10 – 12 years postmortem. However, for some bone samples the PP16 system was not able to amplify many loci using the standard 32 cycle PCR protocol. In most of the samples that failed testing using the PP16 system it has been shown that they contained less than 100 pg of DNA, as analyzed by the Applied Biosystems Quantifiler system, as well as significant levels of PCR inhibitors. A number of studies have shown that increasing the number of PCR cycles can yield more DNA STR data. With this increase in sensitivity it has also been shown that there is an increased risk of producing artifactual peaks, peak imbalances, and there is an increased risk of contamination. As a first step to try and obtain more STR data from the DNA extracts, with low levels of DNA recovered, the PP16 system was tested at increased cycles and optimized for low copy analysis on these extracts. To study the effects of LCN analysis on the PP16 system various DNA samples, containing between 20 pg and 500 pg of DNA, were amplified between 32 and 36 cycles. The effects of the various levels of DNA were compared for the occurrence of stutters, artifacts, heterozygote balance and allelic drop out. The results from this study are compared to previously published data for the PP16 system as well as that of the Applied Biosystems AmpFlSTR® SGM Plus kit. LCN, STR, DNA B7 Optimization of the Promega PowerPlex® 16 System for Testing of Bone Samples With Low Levels of DNA Stojko Vidovic, PhD*, Daniel Vanek, PhD, Zoran Obradovic, MD, Bozana Filipovic, Danijela Music, Jon M. Davoren, MS, and Tony Donlon, BS, International Commission on Missing Persons, Alipasina 45a, Sarajevo, 71000, Bosnia and Herzegovina After attending this presentation, attendees will be shown the results of the optimization of the Promega PowerPlex® 16 system for use on skeletal samples. This presentation will impact the forensic community and/or humanity by demonstrating the significant benefit for anyone performing DNA-STR analysis of bone samples. The International Commission on Missing Persons has been given the task of identification of missing persons from throughout the former Yugoslavia. This identification process is challenging primarily because of the large number of missing persons, the relatively long length of time that has passed since persons have gone missing, the lack of medical records, and the large number of sets of co-mingled remains. The process of reassembling bodies by DNA STR testing is often difficult because many of the less dense bones contain much lower levels of DNA. The low levels of DNA in many bones such as ribs, vertebrae, and pieces of skull bones makes the STR testing very difficult. In order to optimize DNA STR testing procedure for bone samples with relatively little DNA researchers have investigated the effects of altering a number of parameters in the Promega PowerPlex® 16 system (PP16). The PP16 system has been optimized by altering the amount of primers added to the PCR reaction, increased amounts of taq, increasing the length of the extension cycle and concentration of the final amplified products. As the DNA isolated from small amounts of bones is well below the recommended amount of DNA for the PP16 system all optimizations were * Presenting Author validated to ensure that quality of the results. The optimizations of the PP16 system have been shown to increase the success rate of the testing from 0% using the PCR protocol in the PP16 manual to 30 – 40% with the modified protocol. DNA, STR, PP16 B8 The Relationship Between DNA Quantity and the Quality of the DNA STR Profile for DNA Extracted From 10- 13-Year-Old Bone Samples Rijad Konjhodzic, BS*, Jon Davoren, MS, Ana Milos, BS, Arijana Selmanovic, BS, Lejla Smajlovic, BS, Edina Omerovic, Ljiljana Jukic, BS, and Adnan Rizvic, International Commission on Missing Persons, Alipasina 45a, Sarajevo, 71000, Bosnia and Herzegovina After attending this presentation, attendees will be shown results obtained while working with 10 - 13-year-old bone samples. This presentation will impact the forensic community and/or humanity by assisting people working on DNA STR analysis of degraded samples. The International Organization of Missing Persons (ICMP) has been tasked with the identification of approximately 30,000 missing persons. The ICMP identification system has been implemented using deoxyribonucleic acid simple tandem repeat (DNA STR) testing of family reference samples and bone samples. To ensure the accuracy of the identification process the minimum acceptable posterior odds for matching of reference and bone samples has been set at 99.95%. Unfortunately for many cases there are few family members alive to use as reference samples and therefore achieving a posterior odds of 99.95% requires a large number of STRs. The ICMP uses the Promega PowerPlex® 16 system (PP16) as the primary kit for STR analysis. With the ICMP’s extraction procedure the success rate for reporting of 12 or more loci from the first run is approximately 88%. Unsuccessfully tested samples, however, represent a large number of missing, due to the fact that the overall number of victims is so high. Samples that fail the initial testing are quantified using the Applied Biosystems Quantifiler™ system. The Quantifiler™ kit uses Real-time PCR technology to obtain both the amount of DNA present in a sample as well as to assess the level of inhibition that occurs during the PCR amplification. Quantification was performed on 200 DNA extracts that had already been tested with the PP16 system. For these 200 samples a range of results were obtained during the PP16 testing. Samples were selected that amplified no alleles while others were chosen that amplified between 1 and 16 loci. A correlation was made between DNA content of an extract and the success rate of the STR testing. Samples with fewer than 16 loci amplified were then retested using adjusted amounts of DNA. This presentation will discuss the range of DNA quantities calculated by the Quantifiler™ system and how this information can serve as a means of improving the success rate of the PP16 testing. It will also show how the relatively inexpensive Quantifiler™ testing can both reduce the cost and improve the results of the more expensive STR testing. Ultimately improving the success rate of the STR testing will increase the number of cases of which the posterior odds are over 99.95% and therefore can be identified. DNA, Real-Time PCR, STR * Presenting Author B9 Biological Stains Collected From Crime Scenes Using FTA® Paper Luis J. Martinez-Gonzalez, MS, F. Fernandez-Rosado, MS, Esther Martinez-Espin, MS, J. Carlos Alvarez, PhD, and Carmen Entrala, PhD, Laboratory of Genetic Identification, Department of Legal Medicine, University of Granada, Granada, 18012, Spain; José A. Lorente, MD, PhD*, University of Granada, Department of Legal Medicine, Faculty of Medicine, Granada, 18002, Spain; and Bruce Budowle, PhD, FBI Laboratory, FBI Academy, Quantico, VA 22135 After attending this presentation, attendees will learn about the utility of FTA® paper to recover and store semen and blood stains (from 1 to 10 microliters) from different surfaces (absorbent, non absorbent), as long as the quantitation and amplification protocols that yield better results with these stains. This presentation will impact the forensic community and/or humanity by showing forensics scientists a new approach to recover biological evidences from crime scenes. Proper collection and preservation of biological evidence recovered from the scene of crime is crucial to facilitate the analysis and interpretation of all analytical results, including DNA typing. FTA® paper is a wellknown and widely use medium to collect and store biological materials before completing DNA or RNA analysis. Its typical applications are basically focused on the generation of databases (clinical and forensic ones), although it has also been used for preservation of other biological materials, as those related to agriculture and forestry. Among the main advantages of FTA® is its ability to preserve biological materials. When specimens are spotted or applied to the FTA® matrix cards, cell membranes and organelles are lysed, and the nucleic acids are released, causing both RNA and DNA to become entrapped in the fibers of the matrix. Therefore, biological samples such as blood or saliva can be preserved at room temperature (without further need to cool or freeze), and to rapidly inactivate organisms including blood borne pathogens, preventing the growth of bacteria and other microorganisms. Finally, it is also important to mention that archived samples are ready for analysis in less than 30 minutes, since the genomic DNA remains bound to the FTA® paper; this purification process is easily amenable for automation. Blood and semen stains ranging between 1 and 10 microliters were spotted and collected from 5 different surfaces (wood, cotton/clothes, tiles, glass, and carpet), after 3 different periods of time: one day, one week, and one month. Two different ways of recovery are being used. The first one is accomplished by moistening the FTA® paper with sterile distilled water and then pressing it against the stain. For the second alternative the stain is moistened with sterile distilled water and then the FTA® paper is applied. Results show that FTA® paper is an ideal medium to collect dry specimens from hard, non-absorbent surfaces, such as wood, tile and glass. It is also good medium to collect cells from absorbent surfaces (carpet, cotton), although the performance depends on the size of the stain (amount in microliters of biological specimen spotted). The best approach is to first moisten the sample, and then apply and press with FTA® paper. From non-absorbent surfaces, positive PCR amplification for autosomal STRs, Y-chromosome and correct allelic assignation have been obtained in all cases using as little as 1 microliter of blood or semen. It should be considered that the volume of 1 microliter used to test FTA®’s abilities is clearly smaller than most of the samples found at the scene of crime, where bigger biological stains will be more easily recovered with FTA®. Among the advantages -already presented for bloodstains at the last AAFS meeting- the first one to mention is that this is an easy procedure to recover samples from the scene, since it is only necessary to add sterile distilled water into the sample and apply the FTA® paper; second, it is possible to store samples at room temperature for a long time; third, the preservation of the original support (the place where the biological fluid 35 was deposited), since there is only a transfer of the cells from, i.e., the carpet or the wood, into the FTA®. Finally, a fourth advantage worth mentioning is the relative homogeneity in collecting and storing different kind of samples that could be achieved by using FTA® in a number of cases. FTA®, Scene of Crime, DNA Analysis B10 Species Identification of Kachuga tecta Using the Cytochrome b Gene James Chun-I Lee, PhD*, Hsing-Mei Hsieh, PhD, Central Police University, 56 Shu-Jen Rd, Kwei-San Taoyuan, 33334, Taiwan; Li-Hung Huang, MS, Li-Chin Tsai, PhD, and Yi-Chen Kuo, MS, Department of Forensic Science, Central Police University, 56 Shu-Jen Road, Kwei-San Taoyuan, 33334, Taiwan, Republic of China; Adrian Linacre, PhD, University of Strathclyde, 204 George Street, Glasgow, G1 1XW, United Kingdom; and Chung-Ting Hsiao, MS, Department of Forensic Science, Central Police University, 56 Shu-Jen Road, Kwei-San Taoyuan, 33334, Taiwan, Republic of China After attending this presentation, attendees will learn species identification of Kachuga tecta by DNA tchnique using the cytochrome b gene. This presentation will impact the forensic community and/or humanity by demonstrating how the Cytochrome b gene is a valuable genetic marker in the species identification. A DNA technique has been established for the identification to species level of tortoises. The test works on the shell of the animal, which is frequently used in the illegal production of ornaments and preparation, were used to identify samples from the species Kachuga tecta. A total of 100 tortoise shell specimens collected from the National Council of Agriculture, Taiwan, were used in this study. Primer pairs were designed to amplify partial DNA fragments coding for cytochrome b within the mitochondrial genome. The DNA data showed that among the 100 samples there were four distinct haplotype DNA sequences, within which there were a total of 90 variable sites. Between haplotypes 1 and 2, there is only 1 nucleotide difference at the nucleotide position 228. Between haplotypes 1 and 3, 64 nucleotide differences were observed; haplotypes 1 and 4, 62 nucleotide differences; and haplotypes 3 and 4, 52 nucleotide differences were observed. All four haplotypes were compared to the DNA sequences held at the GenBank and EMBL databases, the most similar species were Kachuga tecta (haplotype 1 and 2), Morenia ocellata (haplotype 3) and Geoclemys hamiltonii (haplotype 4), and the highest similarity were 99.5%, 99.3%, 89.9% and 99.5% respectively. However as this was only 89.9% homologous of haplotype 3 with Morenia ocellata, it would seem that this haplotype shows only limited homology to a similar species registered currently in these databases. The method established by this study is a further method for the identification of samples protected under CITES and will improve the work for the preservation of the endangered species. Kachuga Tecta, Cytochrome b Gene, DNA Forensics B11 Evaluation of Commercial DNA Extraction Kits Using Bacterial Spores Associated With Problematic Matrices Matthew J. Ducote, PhD*, and James M. Robertson, PhD, FBI-CTFSRU, FBI Academy, Building 12, Quantico, VA 22135; and Douglas L. Anders, PhD, FBI-HMRU, FBI Academy, Laboratory Building, Quantico, VA 22135 After attending this presentation, attendees will retain that commercially available kits can provide high quality DNA from problematic matrices, including soils, foods, drinks, and plants. 36 Although the kits tested are intended for microbial samples, this presentation will impact the forensic community and/or humanity by demonstrating how they can also be applied to situations in which the extraction of human DNA is required from problematic matrices. Sensitive and specific PCR-based assays are available for detecting a variety of pathogenic microorganisms which may be used as bioterror agents; however, difficulties with these techniques may be encountered if the organisms are associated with materials such as soils, food and drink items, and plants. Because of inefficient separation of biological material from the surrounding matrix, suboptimal cell lysis, and co-purification of PCR-inhibitory substances along with nucleic acids, traditional laboratory methods for isolating DNA may prove ineffective for such samples. In cases where the release of a biological threat agent results in amounts of potentially hazardous microorganisms higher than those found in the environment, it is essential that high quality DNA can be extracted and used for PCR-based detection and identification. Commercially available DNA extraction kits are designed to efficiently release nucleic acids from bacteria and other cell types typically found in the soil, as well as to remove or neutralize substances which may inhibit downstream applications such as PCR. In this study, several kits have been evaluated with respect to reproducibility and quality of the end product as obtained from a variety of problematic environmental matrices. Loam, clay, and sand samples were spiked with Bacillus cereus spores (a surrogate for B. anthracis), allowed to incubate for various times, and then subjected to DNA extraction using the kits. The various soil types provided differing starting amounts of PCR-inhibitory humic and fulvic acids, as well as different levels of physical attachment of the surrogate cells to the matrix, and therefore different types of challenges to the materials and reagents included in each kit. A variety of food and drink items as well as leaf surfaces were also tested as materials from which to extract bacterial DNA, and represent examples of other problematic matrices that may be encountered in forensic scenarios. Real-time PCR amplification of a 145 base pair region from the B. cereus genome was used to determine the quantity of B. cereus DNA obtained from the spiked samples, as well as the effectiveness of the various kits at procuring high quality DNA free of PCR inhibitors. To optimize the kits for use in the forensic laboratory, the manufacturers’ protocols were modified as necessary, including the addition of an exogenous reagent that has been shown to effectively remove PCR inhibitors. The results of this study will be made available to the forensics community so that researchers and investigators may choose appropriate methods for isolating nucleic acids from different types of biological material incorporated into problematic matrices. DNA Extraction Kits, Bacterial Spores, Problematic Matrices B12 DNA Detergent: A Novel Technique to Remove PCR Inhibitors From Soil-Derived DNA R. Lazarin, BS, R. Wingo, PhD, T. W. Robison, PhD, J. M. Dunbar, PhD, and P.C. Stark, PhD, Chemistry Division, Applied Chemical Technology Group, Los Alamos National Laboratory, MS J964, Los Alamos, NM 87544; James M. Robertson, PhD*, FBI-CTFSRU, FBI Academy, Building 12, Quantico, VA 22135; and Douglas L. Anders, PhD, and Rebecca L. Walker, PhD, FBI-HMRU, FBI Academy, Laboratory Building, Quantico, VA 22135 After attending this presentation, attendees will know that it is possible to remove PCR inhibitors from environmental samples in a one-step procedure. This presentation will impact the forensic community and/or humanity by demonstrating a simple procedure was developed for removal of humic and fulvic acids from forensic samples. Forensic scientists will be given a protocol that can be used to remove humic acids from environmental samples. * Presenting Author Procedures for purification of DNA extracted from bacteria in environmental samples are time-consuming and require expertise by the analyst. The extraction procedures must be complex, because environmental samples are contaminated with humic and fulvic acid substances, which often co-purify with the DNA. If the humic and fulvic acids are not removed from the DNA, they will prevent analysis by the PCR because the dissociation and enzymatic steps are inhibited. In addition, the compounds can influence the results of down stream procedures that utilize fluorescence detection, such as real-time PCR. Commercial kits are available for extraction of DNA from soil samples, but the yield and quality of the DNA is often poor. An alternative to the commercial kits has been developed that has the potential to allow a high number of samples to be processed in a day. The new procedure uses a compound (hereafter referred to as PCR inhibitor remover, PIR) that is added to the crude extracts to initiate an immediate, selective reaction with humic acids and other PCR inhibitory materials. After mixing with the PIR compound, the DNA was precipitated, dissolved in buffer, and a portion tested in the PCR. To identify effective PIR compounds, the approach taken was to examine compounds known to be reactive with carbonyl groups because these moieties are highly present in humic acids and could act to sequester Mg2+ and form covalent bonds with primary amines of the polymerase. The PIR compounds were tested on known humic acid samples for their humic acid-removal capabilities by both fluorescence spectroscopy and PCR compatibility. A popular commercial kit was tested with the humic acid-containing samples in parallel experiments, and the results were compared with those obtained with the PIR compounds. Thiamine, pyridoxamine (PDA), and phenylthiazolium bromide have been identified as promising PIR compounds. These compounds out perform the commercial kit by 10 to 100 fold, improving the PCR amplification 10-10,000 fold over untreated samples. PDA can alleviate PCR inhibition at concentrations exceeding natural levels of humic acids and was selected as the best PIR compound. To utilize PDA in a one-step reaction, it can be tethered to glass beads. An alternative approach is to use serial passage through 5mm and 0.2mm pore size filters to remove the large particles and aggregates of humic acids. A simple protocol will be presented for PCR-ready DNA from soil extracts using the PIR compound and filtration. Exploiting the fact that dilution of extracts often mitigates inhibition effects, it was found that the DNA extract without the PIR compound required up to a 1:10,000 dilution to obtain strong PCR products. In contrast, with the PIR compound, the DNA extract required only a 1:10 dilution to support the PCR. The purification procedure could be completed in less than 3 minutes. Soil, Nucleic Acid Purification, Humic Acids B13 Comparison of DNA Storage Methods Keri L. Smith*, San Jose State University, One Washington Square, San Jose, CA 95192; Steven Lee, PhD, San Jose State University, One Washington Square, San Jose, CA 95192 After attending this presentation, attendees will understand the best methods of DNA storage. This presentation will impact the forensic community and/or humanity by enabling the forensic laboratories to store their samples in such a manner that they are assured that there will be no DNA retention to the plastic. DNA storage is a critical issue in forensic, epidemiological, clinical and genetic database laboratories. 1In forensic DNA laboratories, there is always the possibility that cases may be re-opened and samples may need to be re-tested. This is especially important when the amount of DNA is limited. In addition to sample quantity, intrinsic differences in sample types resulting in differences in quality, and extrinsic differences in the storage buffers (e.g., ionic strength), tube surface type, exposure to UV and temperature of storage may lead to differences in the ability to recover and re-test the sample. * Presenting Author Laboratories utilize different methods to extract DNA depending on the sample type. The composition of the final solvent in the different methods and the inherent properties of the casework samples may impart differences in ionic strength. Casework DNA samples are usually dissolved in TE-4 and kept in plastic tubes. The commercial tubes that are utilized to store the DNA come from different sources and are composed of different types of plastics. It has been observed that DNA can bind to polypropylene tube surfaces and these surfaces cause the DNA to denature.2 Alternate polymers (polyallomer vs polypropylene) appear to reduce the retention of DNA.2 Non ionic detergents have been found to be effective in preventing DNA adhesion at concentrations that do not inhibit PCR.3 In addition some of the tubes may contain nucleases and chemical contaminants that may digest and/or denature the DNA.2 In this study, a comparison of storage of DNA samples at varying concentrations, in different buffers over varying amounts of time, at different temperatures in different tube types, will be performed. Utilization of the most efficient storage method (buffer, tube, and temperature) may prove critical in the ability re-test samples. Inter- and intra-lot tube variation will first be evaluated using control DNA. Glass tubes will be used as a control. Any variation in storage temperatures and humidity will be evaluated and monitored using NIST certified digital thermometers. Tubes containing replicate samples at 0.5, 1.0 and 10ng of DNA in 30ul volume for each time point and each temperature and each tube type will be stored in the dark and covered in aluminum foil to avoid exposure to UV. Samples will be stored at –20°C, 4°C, and room temperature. Aliquots will be analyzed at the start, 1 day, 1 week, 1 month, 3 months, 6 months and 1 year (a 3 year and 10 year time point set will also be made). Samples will be in TE storage buffer and Chelex storage buffer. Quantification will be performed by UV spectrophotometry and a subset will be analyzed using agarose gel electrophoresis. These values will be compared to the original sample values to determine if there has been any loss/retention of the DNA sample based on storage method or length of time. Data in triplicate for each sample type, storage tube and temperature will be analyzed for standard deviation and coefficient of variance. In addition, amplification of STRs from a subset of the samples will be performed. References: 1. Steinberg K, et al., “DNA banking for epidemiologic studies: a review of current practices.” Epidemiology 14, no 2 (2003): 254-5. 2. Gaillard C. and Strauss F. “Eliminating DNA loss and denaturation during storage in plastic microtubes.” Technical Tips Online (1998) 3. Rensen G. Buoncristiani M. and Orrego C. “Assessment of DNA retention on plastic surfaces of commercially available microcentrifuge tubes.” (2002): available from http://www.promega.com/geneticidproc/ussymp13proc/abstracts/73_buon cristiani.pdf DNA, Storage, Polypropylene B14 Mitochondrial DNA-Based Identification of Forensically Important Sarcophagidae and Calliphoridae (Diptera) in Hawaii Bethany G. Fox*, 373 Awakea Road, Kailua, HI 96734; and Ellen Shimakawa, PhD, Chaminade University of Honolulu, 3140 Waialae Avenue, Honolulu, HI 96816 After attending this presentation, attendees will learn about how to identify different species with mitochondrial DNA sequencing, and how important this is to forensic entomologist in order to determining the postmortem interval correctly. This presentation will impact the forensic community and/or humanity by giving them samples of Sarcophagidae and Calliphoridae 37 species from Hawaii. The mitochondrial DNA sequences will help law enforcement in determining postmortem interval. The poster will present partial mitochondrial DNA sequence analysis from DNA isolated from larvae and adult flies of the Sarcophagidae and Calliphoridae families in Hawaii. The mitochondrial DNA sequences from Hawaii are similar but not identical to flies from other locations around the world. Organic extraction such as CTAB, Pure Gene, and other methods were used to isolate amplifiable mitochondrial DNA, Centricon YM-100 filters were used to further purify the template. The Sarcophaga ruficornis DNA extraction sample contained a concentration of .515 g/ l times 50 l= 25.75 g and the Chrysomya megacephala DNA extraction sample contained a concentration of 1.31 g/ l times 100 l= 131 g. The samples were diluted down to a concentration of 10ng/ l, and between 30 and 70 ng templates were used in each PCR reaction. The pair of primers used were TY-J-1460 and CI-N-1840. The Annealing temperature used in the thermocycler was at a set temperature of 41ºC for 30 complete cycles. Amplification product was verified by a 1% agarose gel electrophoresis containing Ethidiumbromide stain, and yields estimated for subsequent sequencing with the DNA molecular weight marker XIV (100-1500bp). Amplification was successful in all of the larvae and adult fly samples. The samples were sent to the Biotechnology Core Facility at the University of Hawaii in Manoa for sequencing in an Applied Biosystems 377XL DNA Sequencer. The sequences analysis was based on a region between 400 base pairs of the gene for cytochrome oxidase subunit one (CO1). The sequences for the Sarcophaga ruficornis and the Chrysomya megacephala were then compared in an online BLAST search engine of the National Center for Biotechnology Information’s Genbank. The results from the sequence comparison for the Chrysomya megacephala in Hawai’i were similar but not identical to the Chrysomya megacephala from between Lae and Bulolo, Papua New Guinea (Wells and Sperling, 2000) (Calliphoridae, GenBank accession AF295551). The results from the sequence comparison for the Sarcophaga ruficornis in Hawaii did not result in a close sequence similarity with the Sarcophaga ruficornis Genbank entry (AF259511) but instead it closely resembled a Sarcophaga africa sequence from Berkeley, California, a related species (Wells, Pape, and Sperling, 2000) (Sarcophagidae, GenBank accession AF259508). The Sarcophaga ruficornis sequence was also found to be similar to Sarcophaga crassipalpis from Berkeley, California (Wells, Pape, and Sperling 2000) (Sarcophagidae, GenBank accession AF259510). The conclusions of this research resulted in isolation of amplifiable DNA for larvae and adult flies in Hawaii. Mitochondrial DNA sequences from these species in Hawai’i are similar but not identical to the fly species from other regions around the world. This research also shows evolutionary difference between the two specific species in different regions. Being able to identify the Calliphoridae and Sarcophagidae families more easily will help the forensic community in determining the postmortem interval in Hawaii. This work was supported by USDE grant number P217A030070. Mitochondrial DNA, Sarcophagidae or Calliphoridae, Species Identification B15 A Method for Determining the Age of a Bloodstain Kristin N. Nestor, BS*, Stacey Anderson, BS, Brandi Howard, BS, Regina Trott, Gerry Hobbs, and Clifton Bishop, PhD, West Virginia University, Department of Statistics, PO Box 6057, Morgantown, WV 26506-6057 The goal of this presentation is to show how RNA can also be used in the forensic community as an indicator for the age of a bloodstain. As a result, the age of a bloodstain could act as information in relevance to the time a crime occured. In addition, this information could also be useful for the exclusion or inclusion of a suspect. This presentation will impact the forensic community and/or humanity by informing the forensic science community about new research 38 and technology that can be useful in investigations. In addition, this presentation may bring further interest to this area of study. DNA allows for the unambiguous identification of the person from whom a biological sample was derived but provides little information about when the sample was deposited. This information only indicates that the biological sample was deposited at the crime scene prior to the collection of evidence. The ability to determine the age of a biological sample would greatly benefit the forensic science community, providing a temporal linkage of biological evidence to the time a crime was committed. Conversely, if the sample were deposited at a different time, then value resources might not be wasted pursuing an innocent person. The authors have used real-time reverse transcriptase PCR (RT-PCR) to show that the ratio between different types of RNA (mRNA versus rRNA) changes over time in a linear fashion when dried human blood was examined over the course of 180 days. One study focused on the comparison of individuals from three ethnic groupings: European-American, Asian-American, and African-American. Additional studies demonstrated how environmental conditions, specifically temperature as well as humidity, changes the rate of RNA decay. Although other approaches have been used in the past to estimate the age of a biological sample, this approach offers the following advantages: enhanced detectability of small samples, simultaneous isolation of DNA and RNA from the same sample, species-specific probes, and an increased window of usefulness. Forensics, Real-Time PCR, RNA Decay B16 Analysis of DNA Mixture Samples: Integration of the Quantifiler™ Real-Time PCR Kits and the AmpFlSTR®Yfiler™ Kit Chien-Wei Chang, PhD*, Julio J. Mulero, PhD, Robert L. Green, BA, and Lori Hennessy, PhD, Applied Biosystems Inc., 850 Lincoln Centre Drive, Foster City, CA 94404 After attending this presentation, attendees will that sensitive, reproducible and reliable methods are available to quantify and genotype mixture DNA samples containing relatively low proportions of male DNA. This presentation will impact the forensic community and/or humanity by demonstrating advances in mixture DNA sample analysis for quantitation and STR analysis. This presentation will discuss the incorporation and utility of the Quantifiler™ Real-Time PCR Kits and the AmpFlSTR®Yfiler™ Kit into the analysis of mixture samples typically encountered in forensic casework. Forensic samples from sexual assault cases frequently contain a mixture of genotypes in which the DNA of the male contributor, present only in a very small amount, needs to be discerned from a high background of female DNA. Physical separation of the two genotypes using the differential extraction method may sometimes be possible. However, the technique is laborious, time-consuming and is limited to certain sample sources. With current autosomal short tandem repeat (STR) typing systems, male DNA in mixture samples can be correctly interpreted only when it comprises 5% or more of the DNA in the mixture. Although the degree of male contribution can be estimated by the analysis of the amelogenin locus or by estimating the quantity of male DNA from the number of spermatozoa detected, genotyping results of mixture samples may require a significant degree of interpretive skill from the analyst. With the aim to improve the ability to obtain useful genotypic information from mixture samples containing relatively low proportions of male DNA, recent advances in DNA analysis of forensic samples have focused on developing sensitive, reproducible and reliable methods to quantify and genotype mixture DNA samples. The AmpFlSTR®Yfiler™ PCR Amplification kit is a STR multiplex assay that co-amplifies 17 Y chromosome STR loci in a single PCR reaction. The authors have evaluated the efficacy of the * Presenting Author AmpFlSTR®Yfiler™ typing system combined with the Quantifiler RealTime PCR Kits to determine the range of ratios of mixture DNA within which positive detection and accurate genotyping of a minor male contributor DNA could be obtained. First, mixtures of purified male and female DNAs were prepared and analyzed with the Quantifiler™ Y and Quantifiler™ Human assays to determine the relative contributions of male and total human genomic DNA in samples, respectively. Mixtures with ratios ranging from 1:1 to 1:8000 (male:female) were then processed with the AmpFlSTR®Yfiler™ kit. The characteristics of Yfiler profiles produced from DNA sample mixtures were studied on the basis of specificity, signal quality and intensity. These experiments indicated that full profiles of minor male contributors were obtained from male-female mixture samples with ratios up to 1:4000. In contrast, the male components were not detectable by standard autosomal STR typing methods. The resulting signal intensity of the Yfiler assays directly correlated with the concentrations of male DNA determined by the Quantifiler™ Y assays. In addition, it was shown that the AmpFlSTR®Yfiler™ typing system allows alleles from multiple male individuals in mixtures sample to be easily determined in the presence of excess female DNA. Results clearly illustrate that the use of the Quantifiler™ Y Human Male DNA Quantification Kit in conjunction with the AmpFlSTR®Yfiler™ Kit allows the detection, quantification and genotype determination of the male DNA component specifically without interference from female DNA. Mixture DNA Sample, STR, Y Chromosome B17 Pitfalls in Crime Scene Investigation: Report of Four Cases Francesco Introna, PhD*, Antonio De Donno, MD, Simona Corrado, MD, and Domenico Urso, MD, Section of Legal Medicine DI.M.I.M.P, University of Bari, P.zza Giulio Cesare, 11, Bari, 11-70124, Italy After attending this presentation, attendees will learn about 4 cases of murder in which avoidable errors committed by the Police scene of crime officers compromised the final result of the investigations This presentation will impact the forensic community and/or humanity by showing the importance of the presence of a team of medical experts in crime scene investigations. This presentation will describe four cases of murder in which avoidable errors committed by the Police scene of crime officers compromised the final result of the investigations. Inspection of the crime scene has a primary role in correct operational set-up and must be meticulously performed by qualified, specifically trained staff. Attention must be paid not only to macroscopically evident signs but also to all elements, however apparently insignificant, present on the scene. In the first case (Court of Bari) a child disappeared from the yard of the apartment block where she lived. Searches were immediately made by the Police Force and continued, unsuccessfully. The only element neglected during the analyses was later to prove decisive: the sudden emergence of flies from a hole in the cellar wall. This had been ignored and the hole filled. A year later the skeletal remains of a child were found under a thin layer of gravel in the apartment block cellar. The Pathologist established a correspondence between the times of disappearance of the child and of death and deposit in the cellar. However, the advanced stage of decomposition made it impossible to establish by autopsy and laboratory investigations whether prior rape had been committed. The fundamental error in this case was the incomplete search for the body in the vicinity of the child’s habitation. In the second case (Court of Lecce), a Mafia murder and summary burial of the body (a “lupara bianca” case), the skeletal remains were committed to the Section of Legal Medicine DI.M.I.M.P for investigation after superficial excavation by the police force of the hole in the ground where the body had been found. Examination revealed a firearm entrance wound * Presenting Author in the skull. No exit wound was apparent, but the bullet had not been retained in the skull. The authors felt obliged to make a second, meticulous excavation of the crime scene using metal detectors. This yielded not only the bullet but also other bone remains that the police officers had failed to recover. Examination of the bullet resulted in identification of the weapon from which it had been fired and hence of the murderer. The error lay in the incomplete excavation of the hole and recovery of the skeletal remains performed by inexpert staff. The third case (Court of Lecce) also regarded a Mafia murder and burial of the body (lupara Bianca), where the confession of a “pentito” (an erstwhile accomplice) had indicated ritual lapidation as the murder method. During the excavation operations, carried out by the undertakers, a vast accidental fracture was caused in the occipital region by a stroke with a spade. This postmortem lesion allowed the defence to claim that all the lesions were postmortem and thus to confute the accusation of murder by lapidation. Close anthropological examination of the injuries then enabled differentiation between the vital lapidation injuries and the postmortem lesion caused during recovery. This confirmed the pentito’s confession and the State Attorney secured a murder sentence. Again the error was in allowing inexpert staff to excavate the hole and recover the skeletal remains. In the last case (Court of Bari), a 7-year-old gypsy disappeared. Searches by the Police Force, with search dogs (later reported to have antidrugs training), were unsuccessful. The girl was assumed to have been kidnapped by a rival clan, but not murdered. After about 4 months, a cadaver in a pre-skeletrization state was found 400 metres from the site of disappearance, at the edge of a rural road. Investigations confirmed a correspondence between the times of disappearance and of death. No objective signs could establish the cause of death or any prior rape, and the case is still unsolved. The error was in allowing unqualified staff to search for the body. In the light of the above cases, the authors emphasize that in crime scene investigations involving cadavers or biological materials, the presence of a team of medical experts in criminology, pathology or forensic anthropology is essential to avoid missing vital clues. Both pathologists and police officers should therefore be selected from among highly qualified staff, and should undergo periodic updating courses. Crime Scene, Investigations, Pitfalls B18 Education of the Forensic DNA Analyst in the 21st Century Joseph E. Warren, BS, MS, PhD*, John V. Planz, BS, MS, PhD, and Arthur J. Eisenberg, BS, MS, PhD, University of North Texas Health Science Center, Department of Pathology and Anatomy, 3500 Camp Bowie Boulevard, Fort Worth, TX 76107 After attending this presentation, attendees will gain knowledge of a graduate program specilaizing in training forensic DNA analyst and technical leaders. This presentation will impact the forensic community and/or humanity by informing the forensic community the existence of this program and its potential to assist the forensic community in the challenges of 21st. Century DNA testing. It is hoped that a dialouge will be started with Laboratory Directors and DNA Technical Leaders as to how the faculty can further design this program to keep it as relevant as possible in order to meet the current and future needs in the field. Accreditation of Forensic DNA testing laboratories has now become standard in the field. An important aspect of the accreditation process is the qualifications and training of the DNA analyst. The National Standards has explicit requirements for coursework and training of DNA analysts and Technical Leaders. Furthermore, with the advent of new technologies such as SNP panels, Real-Time PCR, robotic systems, non-human DNA testing, mitochondrial DNA testing, Y chromosome analysis and Whole Genome 39 Amplification individuals who are trained to understand the science, research, development and validation of these systems will soon be required by crime laboratories to implement these technologies. Other disciplines that are now developing including microbial forensics, bioterrorism, genomics and bioinformatics in forensic science, will require individuals with at least some familiarity with these areas. In 2001, the University of North Texas Health Science Center started what was at that time, the first graduate program to offer a Master of Science degree specializing in Forensic Genetics. This program was developed to provide the forensic community with trained personnel, all of who meet and exceed the requirements to be DNA Technical Leaders except for the actual three-year work experience as a DNA analyst in a forensic laboratory. While that is still the primary focus of the program, it is also recognized by the faculty that it must keep abreast of new discoveries and technologies available in genetics and identity testing so as to produce the finest students capable of working in 21st century forensic laboratories. The program itself is two years in length. The first year consists of coursework in molecular biology, biochemistry, ethics, biostatistics, a journal club entitled “Topics in Forensic and Molecular Genetics,” population genetics, immunology and microbiology. During the summer of year one the students take two courses, Molecular Methods in Forensic Genetics and QA/QC for Forensic DNA Laboratories. Second year coursework includes courses in Biological Evidence Evaluation, Genetic Data Analysis, Topics in Forensic and Molecular Genetics, Forensic Anthropology and Expert Testimony in Forensic Science. Four electives are also offered, Bloodstain Pattern Analysis, Forensic Hair Comparison, mtDNA Sequencing and Forensic Biology: The History and Science of Human Identity Testing. The last project the students need to complete is a 6-8 weeklong internship at a forensic laboratory. They then are required to present that and publicly defend it as an internship practicum. This presentation will provide more information as to the coursework, research projects, job placements and future projects of this unique program. DNA, Education, Training B19 An Evaluation of the Efficacy of a Large Metropolitan Detective Training Program in Forensic Biology/DNA Concepts and Procedures Utilizing a Hierarchical Formative Evaluation Garry J. Bombard, MS*, Forensic Institute for Research, Science, and Training, 3400 West 111th Street, Suite 116, Chicago, IL 60655 The goal of this presentation is to present to the forensic science community an educational hierarchical formative evaluation model for use in evaluating forensic biology/DNA training to police detectives/ investigators. This presentation will impact the forensic community and/or humanity by providing an example of a public policy analysis conducted by a forensic scientist. The analysis introduces an evaluation model measuring the effectiveness of a forensic biology/DNA training program to police detectives. The model can be further utilized to develop and evaluate the effectiveness of any future forensic science training within the criminal justice community. The presentation is timely and applicable based upon several national agendas. The presentation provides an overview of a public policy analysis. The public policy analysis is a research study examining the efficacy of a collaborative forensic biology/DNA training program to the Chicago Police Department Detective Division. The study establishes a unique model of evaluation derived from the educational field. The study is timely in developing forensic science training programs to criminal justice agencies and 40 mirrors the national agenda for the development and monitoring of training programs. The specific focus is to determine the efficacy of training, evaluating knowledge and understanding of forensic biology/DNA concepts and procedures, of a forensic biology/DNA training program previously presented by the Illinois State Police Forensic Science Center at Chicago to the Chicago Police Department Detectives. The study evaluates the retention of information by the training attendees, the ability to utilize the information, and identifies future training needs of the Chicago Police Department Detective personnel. The study develops a model to measure the efficacy of the collaborative training program. The researcher utilizes an educational model, a hierarchical formative evaluation, proposed in 1993. The hierarchical formative evaluation is further developed and operationalized by incorporating a widely accepted educational taxonomy on cognitive thinking skill levels. The resulting evaluation model is a unique approach to measure the efficacy of training at several different levels. The final evaluation model is unique. No published literature documents the incorporation of this taxonomy into the formative evaluation model. A self-administered survey instrument is the primary data source. The survey instrument (questionnaire) was developed from material presented in the collaborative training program. The questionnaire measures the cognitive thinking skill levels for achievement of the material delivered during training. From a local perspective, the benefits of the study determine the current understanding of forensic biology/DNA within the Chicago Police Department Detective Division. The study also determines if the training received from the Illinois State Police was effective and identifies additional specific topic related training. In addition, suggestions for changes/modifications to the current program for new detective classes are recommended. From a national perspective, the research is applicable to other entities within the criminal justice system and other complex systems/organizations. To date, very little emphasis has been placed on the development of training in forensic sciences for police detectives. This lack of training and emphasis in forensic science has been identified through national surveys and studies on police detective training. Currently, there is the start of a national effort, the “Improving Justice Through DNA Initiative,” to develop formal forensic science training programs for all elements of the criminal justice community. Additionally, the research will validate the collaborative training model. The collaborative approach to the development of the training is new to the law enforcement community. The collaborative model demonstrates a successful method to accomplish forensic science education in detective training in the future. This research on the development of the hierarchical formative evaluation model and confirmation of the collaborative training model will significantly contribute to the national agenda. Lastly, the National Institute of Justice (NIJ) is developing a model to measure the effectiveness of training. The evaluation of training will be mandatory for any training project utilizing grant money. The model presented accomplishes this future requirement. Forensic Biology/DNA Training for Detectives/Investigators, Public Policy Analysis, Hierarchical Formative Evaluation B20 Microspectral Analysis of Gemstones and the Applications to Forensic Science Paul Martin, PhD*, CRAIC Technologies, 2400 North Lincoln Avenue, Altadena, CA 91001 After attending this presentation, attendees will learn the techniques used to prepare and analyze gemstones and to understand and interpret the spectral data. Since gemstones are often stolen, this presentation will impact the forensic community and/or humanity by giving the forensic community the * Presenting Author skills to identify and characterize gemstones as criminalists comes across them in their work. The theft of gemstones has occurred since time immemorial and has become quite romanticized. The topic has been the subject of literature and film from “Le Count du Monte Cristo” to “The Pink Panther.” Yet the theft of gemstones is quite a real issue and there are real problem in identifying gems. In addition, because of their value, gems are often counterfeited with less precious materials. It is important for the forensic scientist to be able to confirm the identity of precious stones and their less precious substitutes. The purpose of this paper is to show the techniques used for the forensic analysis of gemstones by UV-visible-NIR microspectroscopy and microfluorometry. These techniques include sample preparation, methods of spectral data acquisition and, of course, spectral analysis and interpretation. The talk will also review the data from several types of gemstones in order to aid the examiner in identifying them in casework. mixed with anticoagulant was also spilled. Automatic hourly recordings of temperature, windspeed and precipitation were obtained for the area during the entire study period. The areas of spilled blood were monitored daily while blood was visible with the naked eye. Thereafter selected areas within the areas where the blood was spilled, were sprayed weekly with luminol and the luminesence documented. Planned study period is 18 months. For an extended time frame, a homicide scene where blood was known to have been spilled in a lava field 18 months previously was revisited with luminol at the onset of the study. Results of this study will show the usefulness of applying luminol at outdoor crime scenes to detect diluted blood, will discuss the different results on different types of soil and discuss how it may aid the crime scene investigator in his/her homicide investigation. Luminol, Outdoor Crime Scene, Criminalistics Microspectrophotometer, Gemstones, UV-Visible-NIR B22 B21 The Outdoor Crime Scene: Influence of Weather and Soil Types on the Detection of Diluted Blood With Luminol Thora S. Steffensen, MD*, University Hospital of Iceland, Department of Pathology, Rannsoknastofa H.I. vid Baronsstig, Reykjavik, IS 108, Iceland; and Omar Palmason, Reykjavik Police Department, Hverfisgata 115, Reykjavik, IS 108, Iceland The goal of this presentation is to demonstrate to the forensic community the usefulness of luminol application at outdoor crime scenes, months and even years after blood was spilled. This presentation will impact the forensic community and/or humanity by showing the usefulness of applying luminol at outdoor crime scenes to detect diluted blood, will discuss the different results on different types of soil and discuss how it may aid the crime scene investigator in his/her homicide investigation. Traditionally luminol has been used as a presumptive reagent to detect blood, both fresh and old. Luminol is capable of detecting blood in dilution up to 1:5.000.000 and it´s usefulness at indoor crime scenes is well known and well documented. Due to the effect of rain or snow, visible blood may quickly disappear at an outdoor crime scene. Little has been documented on the use of luminol to detect diluted blood in outdoor conditions although homicides committed outdoors are not uncommon. This presentation will describe a study performed to investigate the time period that luminol can detect diluted blood outdoors, in exposed areas and to investigate if the type of soil that blood is spilled on, affects the length of time that luminol can detect spilled blood outside. The study was also designed to investigate the possible difference in detection of diluted blood with luminol, between blood spilled in the summer, when the ground is dry, and blood spilled in the winther, when the ground is wet or snow covered. Possible difference in the behavior of human versus pig blood and human blood with and without an added anticoagulant was also investigated for research purposes. A remote rural area with restricted access was chosen for the site of study. The area contained a green area, with both grass and moss, gravel, mixed with sand, and rocks. On the green area as well as the gravel and rock, selected volumes of human blood (obtained from live volunteers) was spilled. The volume ranged from 5ml-1000ml, to simulate anything from insignificant bleeding to a life threatening blood loss. Blood was spilled according to a protocol in the month of August, under sunny and dry conditions, and in February, on top of snow, while raining. As human blood in large volumes is difficult to obtain, a parallell study was run using pig blood for comparison to see if pig blood might be substituted for human blood in further such outdoor studies. Both the human and pig blood was without anticoagulant. For further comparison, a large volume of human blood * Presenting Author Fulfilling the Role of the Forensic Scientist: The New Orleans Police Department as a Case Study and Argument for Greater Emphasis on Actualized Science Anna S. Duggar, MS*, and Rose R. Duryea, MA, New Orleans Police Department Crime Laboratory, 2932 Tulane Avenue, New Orleans, LA 70119 After attending this presentation, attendees will learn the emphasis of active casework and interagency communication over the strict revalidation of accepted techniques. Practical examples from the working New Orleans Police Department structure will be provided as a starting point for efforts in the attendee’s lab or law enforcement agency. Most public sector forensic scientists have to face the choice of research and validation versus casework and communication. Using the NOPD as a model, this presentation will demonstrate the need for scientifically trained personnel to move from the academic emphasis on completeness to the more flexible “triage” emphasis on responsiveness and inter/intra-agency communication. This presentation will impact the forensic community and/or humanity by serving to provide examples of creative solutions to the conflict, and to contining the current discussion of the role of the forensic scientist in criminal investigations. Forensic science is defined as the application of science to questions of law and is, by nature, an applied science, implemented without the benefit of the controlled conditions present in more traditional sciences. Recent accreditation, validation, and quality assurance/quality control efforts aim to regulate science to prevent the acquisition of false data and to make results more reproducible. But this inherently desirable process can go too far if insistence on formal validation protocols in applied forensic labs creates an impediment to a laboratory’s expansion of its capabilities or its ability to meet the needs of the law enforcement community. With the challenges of understaffing and underfunding facing many applied forensic labs, most forensic scientists have to face the choice of research and validation or casework and communication. At the 2004 meeting of the American Academy of Forensic Sciences, a series of presentations in the Criminalistics section addressed the involvement of the forensic scientist at the crime scene and in criminal investigations. Widespread agreement exists that scientists in forensic science must be involved, as non-scientists cannot be expected to produce skillful applications of the scientific method to applied casework. As with any science, experimentation leading to expansion of capabilities and new technologies is vital. Forensic science has an impressive cadre of researchers whose full-time mission is the dissemination of improved technology to the practitioners in the field. But the practitioners have their own challenges to meet: timely casework results—in not only an adjudicative role, but in the underutilized investigative mode—and inter-agency communication must be paramount for the applied forensic scientist to fulfill 41 his or her function. Cumbersome validation studies that prevent or delay the usage of generally accepted techniques can be a significant hindrance to job performance. The New Orleans Police Department is a working model demonstrating the vital need for scientifically trained personnel to move from the academic emphasis on completeness to the more flexible “triage” emphasis on responsiveness and communication. Criminalists at the NOPD Crime Lab have faced the considerable caseload of any forensic scientist in a large city, compounded with drastically low wages and a malnourished laboratory budget. In a situation where responsive casework management, investigative involvement of forensic professionals, and better interagency communication are balanced against the forensic community’s demand for in-house validation and classic structure, it rapidly becomes clear that more creative approaches are necessary. For example, in response to the identification of a lack of communication between branches of law enforcement in New Orleans as one of the primary obstacles facing crimefighting efforts in the city, district attorneys have been invited to COMSTAT meetings, while homicide detectives sit in on the DA’s weekly strategy sessions. Additionally, the constant need for low turnaround times on priority cases has prompted the metamorphosis of the Forensic Light Unit (FLU), a two-man on-call unit, from a reactive latent print unit into a proactive investigative response unit. New techniques are responsibly and effectively put into practice by FLU with appropriate QA/QC protocols, but the predominant emphasis in this new model is the direct application of scientific expertise to questions of law provided by investigators and attorneys, and the conscious commitment of time and energy to maintaining open lines of communication. The success of this new emphasis is evident in both anecdotal and quantifiable levels of satisfaction among investigators. In applying creative problem solving to overwhelming demands on time and resources, the department has definitively demonstrated, as prominent members of the field have suggested, that “doing the job right cannot supplant doing the right job.” The presentation will highlight variations of the typically encountered types of trace evidence that may have greater value given special circumstances and assist in an investigation of an unknown perpetrator or illustrate association with the incident due to the presence of material or form or condition of the samples. Examples to be presented include evidence deposited in bloodstains, beaded spray paint microscopically identified on a roll of duct tape left at the scene which led to a subject, bloodstain patterns of the impression of the weapon (a table steak knife that had been cleaned and returned to a drawer in the kitchen) that demonstrated a garment worn by one of the victims, microscopic examination of a human hair wig used to disguise a perpetrator that led to finding that some imported are made from cadaver hair, the tire and partial license plate impression in snow at the scene of a multiple homicide and variations of fabric damage including an example that helped to substantiate statements of the victim in an acquaintance sexual assault. This presentation will also include some unusual types of trace evidence. Examples include botanical samples such as thorns in a subject’s clothing that were found to be consistent with thorns from a rose bush damaged by the perpetrator who jumped from a second floor window to escape from the scene, sequential layers of material deposited in hollow point projectiles, welding spheres microscopically identified from the debris on the clothing of an 8-year-old kidnapping victim, and air bag evidence such as makeup, fiber samples and starch granules that may assist in determining the driver of a vehicle involved in a fatal accident. If you do, you could say that provided information linking the items to the airbag. The author gratefully acknowledges the permission of Glenn Schubert and the Illinois State Police for permitting the inclusion of photos related to air bag evidence in this presentation. In summary, it should be emphasized that the use of trace evidence as an additional investigative tool may solidify the findings in any given case. Forensic Science, Validation, Communication B24 B23 The Value of Trace Evidence as an Investigative Tool G.M. Yezzo, BS*, Ohio BCI&I, 1560 State Route 56, PO Box 365, London, OH 43140 The goal of this presentation is to provide some case examples of unexpected circumstances or forms of trace evidence that have provided assistance in an investigation. This presentation will impact the forensic community and/or humanity by serving as a reminder of some of the uses of trace evidence as an additional investigative tool that may solidify the findings in case. DNA analyses alone may not resolve all cases that include biological evidence such as semen or blood. Even in cases where these materials are present, depending upon the case facts, trace evidence may assist substantially in the investigation. Most forms of Trace evidence provide only class or comparative information. However, the microscope may be used to identify particles that provide important investigative information. Many of the types of materials discussed in this presentation are documented in available references on trace evidence such as Volumes of The Particle Atlas (McCrone and Delly (1973) and McCrone, Delly and Palenik (1979)), Color Atlas and Manual of Microscopy for Criminalists, Chemists and Conservators (Petraco and Kubic, 2004), Mute Witnesses –Trace Evidence Analysis (Houck, 2001) and Trace Evidence Analysis (Houck, 2003). Taking advantage of the trace evidence may provide crucial investigative information or establish a link between individuals or an individual and a location. The goal of this presentation is to provide some case examples of unexpected circumstances or forms of Trace evidence that have provided assistance in an investigation. 42 Trace Evidence, Investigative Information, Criminalistics Forensic Intelligence - Forensic Science Beyond the Courtroom Rebecca Bucht, BSc*, 309 West 76th Street, #4A, New York, NY 10023; and Oliver Ribaux, PhD, Université de Lausanne, Ecole des Sciences Criminelles, Lausanne-Dorigny, BCH, 1015, Switzerland The goal of this presentation is to introduce a framework for integrating forensic evidence into the criminal investigation and intelligence analysis processes, complete with examples of real life applications and results. This presentation will impact the forensic community and/or humanity by demonstrating how the development of forensic intelligence can improve knowledge of criminal mechanisms through the exploitation of a solid set of real-time data at a tactical, operational and strategic level. Despite evidence of the benefits of applications of forensic case data to the investigative and intelligence world, research and implementation of forensic science continues to focus primarily on its function as proof in courts of law. This not only deprives said investigative and intelligence applications from an added information source, it also renders a huge wealth of forensic information unexploited. Forensic intelligence is in essence the product of logically processing forensic case data. Although the concept of collecting and processing forensic data is by no means novel, modern technology provides databases capable of storing, handling and sorting large amounts of increasingly complex data, allowing these old concepts to be realized more effectively and on a larger scale. The basic inferences made from forensic data include identifying recidivists, individualizing a source based on a trace, individualizing a trace based on a source, linking cases by trace-trace comparisons, profiling of the source of a trace, classifying the source of a trace, listing possible sources of a trace and listing possible relatives of the source of a trace. These inferences are made based on physical and chemical characteristics of samples and standards. * Presenting Author Fingerprint and DNA databases are two examples of where forensic information is used in a wider context, whether it is to identify recidivists, individualize a source based on a trace, individualize a trace based on a source, or link cases. Other forms of traces such as shoeprints, tool marks, and glove prints are less systematically exploited. These traces are abundant on crime scenes, less expensive to process, and can provide significant information. Even though a particular trace cannot necessarily provide a conclusive enough inference to be used as evidence in a court of law, the information it can provide can be indispensable in an intelligence setting. The forensic science community should be more aware of the importance of extending the impact of forensic science beyond the courtroom. Similarly the intelligence community needs to be aware of the kind of information forensic science can provide. Effort also needs to be made to ensure functional channels of communication exist between forensic scientists and investigators, intelligence analysts and other professionals in order to ensure that the forensic case data is properly understood and contextualized. For example, in the case of illicit drug profiling, the chemical data is first interpreted by forensic scientists in order to evaluate links between seizures. This linkage information is then passed on to investigators, and further to criminologists and geo-politicians, who all may be able to combine the forensic data with the data they already base their investigations on. There are many potential benefits in expanding the use of forensic science data in investigative and intelligence scenarios. The input and cooperation of the forensic science community is needed in order to assure a successful integration. Forensic Intelligence, Intelligence Aanalysis, Linkage Blindness B25 1,2-Indanedione: Is it a Useful Fingerprint Reagent? Chris Lennard, PhD*, Australian Federal Police, Forensic Services, GPO Box 401, Canberra, ACT 2601, Australia; Christie WallaceKunkel, BSc, and Claude Roux, PhD, University of Technology, Sydney, Centre for Forensic Science, PO Box 123, Broadway, NSW 2007, Australia; and Milutin Stoilovic, MSc, Australian Federal Police, Forensic Services, GPO Box 401, Canberra, ACT 2601, Australia After attending this presentation, attendees will gain an understanding of the fingerprint reagent 1,2-indanedione: its strengths, weaknesses and the inconsistencies reported over the last 8 years in terms of its value as a fingerprint reagent. Recommendations will be made based on the results of the research performed. This presentation will impact the forensic community and/or humanity by the presentation of the results of this research which will contribute to the body of information already available on the value of 1,2-indanedione as a fingerprint reagent. Conflicting results have been reported to date. This study, under Australian conditions, indicates that 1,2-indanedione is a viable alternative to ninhydrin and DFO for latent fingermark detection on porous surfaces. Ninhydrin is the accepted, routine reagent for the chemical detection of latent fingermarks on porous surfaces such as paper. Ninhydrin reacts with the amino acid component of the latent fingerprint deposit to produce a dark purple product known as Ruhemann’s purple. Secondary treatment of a ninhydrin-developed mark with a zinc or cadmium metal salt produces a coordination complex that is luminescent under certain conditions. Significant enhancement is possible in the luminescence mode. Numerous research projects have been undertaken since the early 1980s to find ninhydrin analogues that offer advantages over ninhydrin itself. In 1995, whilst researching ninhydrin analogues, Jouillé and coworkers, in conjunction with the U.S. Secret Service, synthesised a new type of latent fingerprint visualising agent, 1,2-indanedione (Ramatowski, * Presenting Author 1997). Since then, researchers around the world have conducted research into the capabilities of 1,2-indanedione as a fingerprint reagent. The results have been varied, with conflicting results and different recommendations made. A critical review of the literature regarding 1,2-indanedione was performed. A number of discrepancies exist in the literature as to which formulation and which development procedure produces optimal results. As a result, this project set out to determine the best formulation and development procedure under Australian conditions, encompassing all published recommendations as well as some novel approaches. 1,2-Indanedione formulations were compared with respect to initial color, luminescence, concentration of reagent, acid concentration, and the effect of different carrier solvents. Numerous development conditions were investigated, including a conventional oven, a heat press, humidity, metal salt treatment and liquid nitrogen. A heat press set at 165º C for 10 seconds proved to give the best initial color and the highest luminescence. Secondary metal salt treatment improved initial color and luminescence. The Polilight (Rofin), the VSC2000 (Foster & Freeman), and the Condor Chemical Imaging Macroscope (ChemImage Corp) were used to detect the fingerprints developed with 1,2-indanedione on a variety of high- and low-quality porous surfaces. Overall, very good results were obtained. The optimum formulation for 1,2-indanedione was found to be a 1% (w/v) solution using HFE 7100 as the carrier solvent. 1,2-Indanedione treated fingermarks were best developed using a heat press set at 165º C for 10 seconds, after which they display a bright pink initial color and a high degree of luminescence at room temperature. Field tests were also conducted to compare 1,2-indanedione with DFO and ninhydrin, showing that 1,2-indanedione develops more fingerprints than the conventional reagents and thus should be considered as a viable alternative for the development of fingerprints on porous surfaces. It is obvious that 1,2-indanedione, as a fingerprint technique, is very much dependant on environmental conditions, reagent formulation, and development conditions for optimum results to be obtained. It is for this reason that different research groups around the world have had varying results with respect to the application of this reagent. It has been well established that ninhydrin reacts with amino acids to form Ruhemann’s purple. Very little information on the chemistry of 1,2indanedione exists in the literature and the product formed upon reaction with amino acids has not been confirmed to date. The current research aims to characterize the reaction product using fluorescence spectroscopy, infrared spectroscopy, UV-visible spectroscopy, mass spectroscopy, and nuclear magnetic resonance spectroscopy. Latent Fingermarks, Porous Surfaces, 1,2-Indanedione B26 The Billiard Ball Ricochet Effect and the Garment as an Intermediate Target H. Bulent Uner, PhD*, Istanbul University, The Institute of Forensic Sciences, I.U. Adli Tip Enstitusu Cerrahpasa, Istanbul, 34303, Turkey; and Ismail Cakir, PhD, Council of Forensic Medicine, Minisrty of Turkey, Adli Tip Kurumu Esekapi Cerrahpasa, Istanbul, 34246, Turkey After attending this presentation, attendees will learn the importance of billiard ball effect while determining firing distance. This presentation will impact the forensic community and/or humanity by demonstrating the importance of billiard ball effect while determining firing distance. From the area of dispersed pellets on the target, a rough estimation of range of fire can be made. There are many factors that affect the dispersion of the pellets like length and diameter of the barrel, size of the pellets, degree of the choke, etc. If a shotgun is fired in contact with a body or at close range, pellets hit the body en masse and are scattered a wide area within the body. This 43 ricochet phenomenon has been termed ‘the billiard ball ricochet effect’. Also, this phenomenon can occur when pellets hit the clothing en masse. This intermediate target will cause an increase in dispersion of the shot. The thickness of the clothing, the nature of the fabric and the number of layers of the garment can alter this dispersion of the shot. In determining the range of fire in decomposed and burned bodies where the skin pattern cannot be seen, x-rays revealing shot patterns may be of considerable assistance. However caution is necessary. In these cases, the billiard ball ricochet effect may lead to erroneous conclusions about the range of fire because of the opinion is formed on basis of the pattern of the pellets on x-rays. Wide dispersion of the shots seen on the x-rays may lead to conclusion of a great range of fire than actually occurred The cases presented are good examples of the billiard ball ricochet effect with intermediate targets. In the first case a woman had a contact shotgun wound of the abdomen. On the x-rays, widespread scatter of pellets were seen within the body. In the second case, although the pellets hit the clothing en masse, the size of shot pattern on the body was over a large area. Shotgun, Pellet Distribution, Billiard Ball Ricochet Effect, Range of Fire B27 Who Shot Muhamed Jamal Al-Dura? Is the Boy Still Alive? Maurice Rogev, MB ChB, 11 Zamenhof Street, Tel-Aviv, 64373, Israel; and Nahum Shahaf, Msc*, Natop, Habiluim st 3, Ramat-Gan, 52297, Israel After attending this presentation, attendees will understand the need to be cautious with media reports unless it’s based on accepted forensic science criteria. This presentation will impact the forensic community and/or humanity by demonstrating the inaccurate impression created by the media in their description of the A-Dura episode on the second day of the Palestinian Intifada, emphasises the need for caution in interpreting critical Death news reports published by the media unless its based on accepted forensic science criteria. Details of the Incident: On September 30, 2000, the second day of the Intifada a crowd of Palestinian policemen and civilians, some armed, attacked repeatedly an isolated Israel Defence Force (IDF) outpost situated at a road junction near Netzarim, an Israeli village in Gaza. French and other television groups televised the incident. The event developed initially as a demonstration and turned violent when Palestinian policemen began to shoot at IDF soldiers in the outpost. French television showed a father and his son Muhammed al-Dura hiding behind a concrete barrel. The 12-year-old boy seemed to be killed and his father seemed to be wounded. The French television alleged that A-Dura was killed by IDF fire. The IDF soldiers claimed that they did not see the pair crouched behind the barrel and they didn’t shoot at them. The pictures of the event were shown all over the world and Muhammad a-Dura became the symbol of the Palestinian intifada. Methodology: No autopsy examination of the body was made by the Palestinians, there are no accurate details of any wounds that may have been caused by the gunfire. The body was not identified according to accepted standards and recognized practices. The identity of the body cannot therefore be accepted as that of Mohammed a-Dura the boy A-Dura was clearly not killed by IDF fire. Aerial photographs were taken of the scene that showed the crossroad and the position of the Israeli Defence Force (IDF) outpost. Television footage of scenes of the riot, the boy cowering with his father and of the funeral procession were examined as well as medical notes and still photographs of the boy lying on a mortuary slab. 44 The aerial photographs showed the crossroad, the wall and concrete barrel behind which the boy and his father had hidden, the IDF outpost and the Palestinian outpost called the “Pita” across the road opposite the wall. As the original wall had been destroyed for security reasons, a reconstruction of the immediate area of the wall and the barrel of concrete was made. The barrel had the same diameter of the barrel involved in the incident. Geometric and ballistic criteria were used in the reconstruction The same type of rifles used by the IDF soldiers and the Palestinians were fired directed towards the barrel and the wall at an angle of the alleged fire from the IDF outpost, the “Pita” and from other angles. The television footage and still photographs of the incident and the still photographs of the body were also studied. Medical reports by a Palestinian doctor describing the injuries found on the body were studied in the absence of a complete autopsy protocol. No spent bullets or cartridge cases were available for Ballistic examination. Results and Conclusions: 1. The geometrical and ballistic findings show that the boy was protected from the direction of the Israeli outpost. 2. The spread of the particles of stone caused by the impact of the two bullets next to the boy’s head proves that the fire came from a less oblique direction, consistent with the Palestinian position (named “Pita”). 3. The position of the Palestinians relative to the Israeli outpost raises the possibility that the shooting was deliberate. 4. Part of the bullet holes in the wall were made later artificially. 5. Long cut on the body of the boy described by one of the doctors fit a knife rather than a bullet. 6. The evidence of the doctors shown in the television and in telephone conversations is not consistent with the signs of injury in the photographs of the boy alleged to be that of Mohammed A-Dura in the hospital. This raises the suspicion that this is not the boy A-Dura. 7. According to the evidence of the doctors at the hospital the body of the boy a-Dura reached the hospital some hours before the time of the incident started. Therefore the dead boy presented in the hospital as Muhammad a-Dura cannot be the boy from the Netzarim junction. 8. The signs of injury on the boy’s body at Netzarim were not con sistent with fresh blood. 9. Many manufactured incidents were revealed in the television pictures. These included gunfights in front of the cameras in areas that were hidden from the IDF outpost. The all event around Mohammed al-Dura including many other events seems to be created, including the death of the boy named Muhammad al-Dura. Media Reports, Al-Dura, Forensic Criteria B28 Uncertainty in the Estimated Angles of Impact of Freely Falling Blood Drops Jon A. McGuire, MFS, and Walter F. Rowe, PhD*, Department of Forensic Sciences, The George Washington University, Washington, DC 20052 After attending this presentation, attendees will learn how to determine the confidence limits for the calculated angle of impact of a falling blood drop with a target surface. This preseantation will outline the relevant experimental data for a particular target surface and the statistical treatment of this data. This presentation will impact the forensic community and/or humanity by demonstrating that the uncertainties in the calculated angles of impact of blood drops with target surfaces are considerably smaller than standard texts on bloodstain pattern analysis claim. * Presenting Author The goal of this presentation is to provide crime reconstructionists with a validation of the Balthazard formula for the calculation of the angle of impact of a blood drop with a target surface. This presentation also will also provide estimates for the uncertainty in a calculated angle of impact for ranges of impact ranging from 10 to 90 degrees. Crime scene reconstructionists and bloodstain pattern analysts frequently use the Balthazard formula for calculating the angle of impact of a freely falling blood drop with a target surface from the dimensions of the resulting blood spot. Blood drops impacting a flat surface at an angle 0 produce an elongated blood spot having length L and width W. The angle 0 is then given by the equation: Approximate Angle of Impact 90 80 70 60 50 40 30 20 10 0 = arcsin (W/L) In 1993 the U.S. Supreme Court set out a number of criteria for the admission of scientific evidence; among them is the known or potential error of a scientific technique. Books on bloodstain pattern analysis rarely discuss the uncertainties in the calculated angle of impact. Bevel and Gardner’s widely used text Bloodstain Pattern Analysis suggests that the calculated angles are accurate to within five to seven degrees. This research was undertaken to determine the 95 and 99% confidence limits for the estimated angles of impact of blood drops. Fifteen microliter drops of human blood were allowed to fall ten and thirty-six inches onto the uncoated surface of white poster board with impact angles of approximately 10, 20, 30, 40, 50, 60, 70, 80 and 90 degrees. Fifteen drops of blood were dropped at each angle of impact and each distance of fall. Fifteen microliters was found to be the smallest volume of blood that would fall freely from the disposable tip of a Pipetman pipetter. Two different distances of fall were used to determine if the estimated angles of impact showed any dependence on the distance of fall. Theoretically there should be no such dependence. The untreated surface of the white poster board was chosen as the target surface to reduce the flow of the blood drops after impact. The target surface was held in a homemade device made from Plexiglas and wooden dowel rods. Slots were cut in the Plexiglas at angles ranging from 10 degrees to 90 degrees; the angles of the slots were measured after they were cut. The lengths and widths of the blood spots were measured with a Cen-Tech 4 inch digital caliper. Three of the blood spots produced at an angle of impact of 80 degrees and a thirty-six inch distance of fall were discarded because their widths were greater than their lengths. The measured lengths and widths of the blood spots were used to calculate the angle of impact using the Balthazard formula. The means and standard deviations of the calculated angles of impact were determined for each angle of impact and each distance of fall. Analysis of variance (ANOVA) was performed on the calculated angles of impact. For a ten inch distance of fall an F value of 1187 was obtained, while for a thirty-six inch distance of fall an F value of 1288 was obtained. These results show that the Balthazard formula is statistically significant at the 99.5% level. More importantly, because these two F values exceed the critical F values for the 99.5% confidence level by more than a factor of four, the Balthazard formula is shown to be a satisfactory predictive tool. The standard deviation of the calculated angle of impact was found to increase with the angle of impact, in agreement with previously published work. The confidence range (the difference between the upper and lower confidence limits) for the calculated angles of impact were determined at the 99% confidence level using: where t is Student’s t, s is the standard deviation and n is the number of data points used to calculate s. The table below shows the confidence ranges for the angles of impact from 10 to 90 degrees for both distances of fall. * Presenting Author Distance of Fall = 10 Inches 6.410283 11.2365 3.365768 2.463193 2.148248 1.686314 1.136359 0.795556 1.46632 Distance of Fall = 36 Inches 8.255688 7.701132 4.722746 2.667552 1.754003 2.435882 1.993076 1.59836 0.710401 Up to an angle of impact of 60 degrees the uncertainty in the calculated angle of impact is less than 3 degrees, substantially better than the uncertainty claimed by Bevel and Gardner. Bloodstain Pattern Analysis, Crime Scene Reconstruction, Criminalistics B29 The Detection and Enhancement of Latent Fingermarks Using Infrared Chemical Imaging Mark Tahtouh, BSc, University of Technology, Sydney, Centre for Forensic Science, PO Box 123, Broadway, NSW 2007, Australia; Chris Lennard, PhD*, Australian Federal Police, Forensic Services, GPO Box 401, Canberra, ACT 2601, Australia; and John Kalman, PhD, Claude Roux, PhD, and Brian Reedy, PhD, University of Technology, Sydney, Centre for Forensic Science, PO Box 123, Broadway, NSW 2007, Australia After attending this presentation, attendees will gain knowledge of the capabilities of a novel technique (infrared chemical imaging) for imaging latent fingermarks on difficult surfaces. With technology continuously evolving, this presentation will impact the forensic community and/or humanity by exploring new developments that may prove to be superior to techniques currently in use in forensic laboratories. Infrared Chemical Imaging has been demonstrated to be effective for detecting fingermarks on difficult substrates. This poster will present the results of a study that began in 2003 and continues to date. During this study the use of a new technique, Fourier transform infrared (FTIR) chemical imaging, has been demonstrated for the enhancement of latent fingermarks on a number of surfaces. All fingerprint detection techniques aim to create contrast between the ridge details of a latent fingermark and the background on which it is located. The majority of current methods rely on this contrast to be in the visible part of the electromagnetic spectrum, and thus encounter problems when background interferences such as printed images or patterns are present. Some of these problems may be overcome using traditional visible or fluorescence imaging techniques (e.g., alternate forensic light sources with appropriate barrier filters) or visible and fluorescence chemical (hyperspectral) imaging techniques. However, due to the very broad, overlapping bands that make up all visible absorbance and fluorescence spectra, it is not always possible to obtain acceptable fingerprint images even using chemical imaging techniques in this region of the electromagnetic spectrum. The vibrational spectra (infrared or Raman) of most carbon compounds consist of large numbers (often more than ten) narrow, wellresolved bands that represent vibrational modes of discrete functional groups in these molecules. As a means of identifying and discriminating between different molecules, vibrational spectra are thus far more powerful than UV-visible methods, which consist of very broad, overlapping bands. To date, this inherent feature of infrared (and Raman) spectra has not been utilized in the forensic visualization of fingermarks. This is because that, 45 until recently, infrared and Raman techniques provided no spatial information on a sample. Spatially resolved chemical information, however, is now accessible with the development of infrared chemical imaging. FTIR chemical imaging employs a focal plane array (FPA) detector that can be thought of as a large number (thousands) of discrete detectors (or pixels) laid out in a grid pattern (Digilab, Nicolet, Bruker instruments). The instrument collects images that may consist of thousands of pixels, with a spectrum at each pixel. In this way thousands of mid-infrared spectra are simultaneously collected from a sample while maintaining spatial information. By far the most common type of FPA detector used for mid-infrared chemical imaging is a 64 × 64 pixel mercury cadmium telluride (MCT) focal-plane array detector. The detector used in this study was a Digilab Lancer 64 x 64 MCT detector. This detector collects 4096 infrared spectra simultaneously into a file that can be thought of as a datacube. This three dimensional datacube (x x y x wavenumber) can be visualized as a collection of images of the sample, with one image for each wavenumber resolution unit. This poster describes the application of infrared chemical imaging to the visualization of fingermarks, with and without cyanoacrylate fuming, and explores the future possibilities of this technique. Images of untreated fingermarks on glass substrates with excellent ridge detail were acquired using infrared chemical imaging. High quality fingermarks on glass substrates were also developed using ethyl cyanoacrylate (super glue) fuming and subsequent infrared chemical imaging. This new method allows the collection of images from backgrounds that traditionally pose problems for current fingerprint detection methods. The background may, for example, be highly colored, have a complex pattern, or possess other pattern or image characteristics that make it difficult to separate fingerprint ridges using traditional optical or luminescent visualization. One background that has proven to be a challenging surface for the development of latent fingermarks is the Australian polymer banknote. To demonstrate the power and applicability of infrared chemical imaging, latent fingermarks fumed with ethyl cyanoacrylate were successfully imaged from Australian polymer banknotes. This imaging technique has shown enormous potential for the detection and enhancement of latent fingermarks on a range of surfaces. This work is currently being extended so that the true capabilities of infrared chemical imaging may be realized. It is important to stress that this new method does not aim to replace any of the currently used fingerprint enhancement techniques, but rather aims to provide the forensic scientist with a tool that may prove useful on surfaces where conventional techniques fall short. Infrared Chemical Imaging, Latent Fingermarks, Hyperspectral Imaging B30 Development of a Rubber Stamp for Fingerprint Research Christie Wallace-Kunkel, BSc, and Claude Roux, PhD, University of Technology, Sydney, Centre for Forensic Science, PO Box 123, Broadway, NSW 2007, Australia; Chris Lennard, PhD*, and Milutin Stoilovic, MSc, Australian Federal Police, Forensic Services, GPO Box 401, Canberra, ACT 2601, Australia; and Philip Doble, PhD, University of Technology, Sydney, Centre for Forensic Science, PO Box 123, Broadway, NSW 2007, Australia This presentation will impact the forensic community and/or humanity by demonstrating the use of the rubber stamp for fingerprint detection research will allow fingerprint reagents to be directly and accurately compared not only within one fingerprint research project but across different research projects. Significant applications for quality assurance purposes are also envisaged. Latent fingermarks produced for research purposes are generally deposited by the same donor(s) at various times throughout the course of the project. The problem with this approach is that each fingermark deposited will differ in chemical composition and pressure applied due to environmental and physiological factors. Thus the results cannot be compared directly, as the composition of each latent mark on the support surfaces will vary to some extent. The purpose of this research was to develop a rubber stamping process for the production of ‘standard’ latent fingermarks for the evaluation of fingerprint reagents, ensuring that the same quality print would be deposited with each application, thus allowing direct comparison not only within one research project but across various research projects. The development of the rubber stamp involved three processes: 1) The development and manufacture of a rubber stamp. Inked thumbprints were obtained, scanned onto a computer, and sent to a rubber stamp making company. The stamps were ready within 48 hours. 2) The formulation of a ‘standard’ amino acid solution and the determination of a method of application. The formulation of the solution was determined by investigating the concentration of amino acids in eccrine secretions (human sweat). Inkless stamp pads were obtained to which the amino acid solution was added. 3) Method validation as well as transfer and persistence studies. This involved an investigation of the variations in amino acid concentration between stamp applications. A high performance liquid chromatography (HPLC) method was developed and validated for the detection of amino acids in a water-based solution and in deposited marks. Using high performance liquid chromatography, it was found that the amino acids did not persist on the rubber stamp, nor did the concentration of the amino acids vary over time to any significant extent. This means that each impression laid by the stamp was qualitatively and quantitatively constant — this was the desired outcome for the use of the procedure as a comparative research tool. The whole process enabled a rubber stamp to be developed and used for the application of ‘standard’ latent fingerprint samples to be used for research purposes. Through this process, fingerprint reagents can be directly compared against a uniform ‘standard’. Application of this method in the study of amino acid based fingerprint reagents removes a large number of variables associated with the use of latent fingermarks deposited by a range of randomly-selected donors. A similar strategy could also be used for the development of proficiency tests and quality control checks for the evaluation of latent fingermark detection protocols. To date, no such tests are available. The use of a rubber stamp and an amino acid solution of known concentration cannot replace natural fingermarks, but should be considered as an option for the objective evaluation of an amino acid based fingerprint detection technique. While this option may appear simplistic, it goes some way towards reducing the variables that typically impact on fingerprint detection studies. Latent Fingermarks, Rubber Stamp, Quality Control After attending this presentation, attendees should consider the use of a rubber stamp and an amino acid solution as a method for producing standard latent fingermarks for the evaluation of amino acid specific fingerprint reagents. 46 * Presenting Author B31 Chemical Processes Related to the Development of Latent Blood Fingerprints Melinda K. Lux, BS*, Marshall University Forensic Science Graduate Program, Forensic Science Center, 1401 Forensic Science Drive, Huntington, WV 25701; and Steven P. Kasper, Florida Department of Law Enforcement, Fort Myers Regional Operations Center, 4700 Terminal Drive, Suite 1, Fort Myers, FL 33907 There are many different chemical methods of developing latent blood fingerprints. The goal of this presentation is to address what is the best way to develop these blood fingerprints, especially when those blood fingerprints are found on dark surfaces at a crime scene. This presentation will impact the forensic community and/or humanity by demonstrating how the use of Ninhydrin, Amido Black, and Coomassie Blue are not feasible in these cases where blood is found on dark surfaces because they stain the fingerprint ridges a purple or dark blue color that is not visible on these dark surfaces. The solution may be a different color stain or one that fluoresces the blood print long enough to photograph it. Latent blood fingerprints found on dark surfaces at a crime scene are not useful when developed with Ninhydrin, Amido Black and Coomassie Blue. These three examples are common techniques used on blood prints that stain the fingerprint ridges purple or dark blue which may not be visible on many dark surfaces. The solution to visualize these prints may be a different color stain or one that fluoresces a blood print long enough to photograph it. The chemical processes used in this study are ABTS, Merbromin, IND, Leucomalachite Green, Fluorescein, DFO, and Leucocrystal Violet. The fluorescent nature of many of these processes may assist in visualizing these latent blood fingerprints. Latent, Blood, Fingerprint B32 Evaluation and Comparison of the Electrostatic Detection Apparatus and the Electrostatic Dust Print Lifter on the Development of Footwear Impressions on Paper Christine L. Craig, MS*, Seminole County Sheriff’s Office, 100 Bush Boulevard, Sanford, FL 32733; Breanne Hornsby, BS, 12124 Knights Krossing Circle, #306, Orlando, FL 32817; and Matthew Riles, BS, 1823 Van Pelt Road, Sebring, FL 33870 After attending this presentation, attendees will learn that the Electrostatic Detection Apparatus (ESDA) can be used to develop indented footwear impressions on paper; the attendee will learn the ESDA can be used in conjunction with the Electrostatic Dust Print lifter to obtain footwear impressions from paper; the attendee will learn which technique (the ESDA or the Electrostatic Dust Print Lifter) should be used to obtain the highest quality footwear impression; the attendee will also learn that if both techniques are used what order they should be used in to obtain the best footwear impression; and the attendee will also learn the ESDA will obtain footwear impressions on the top sheet of paper if a stack of papers are stepped on and the Electrostatic Dust Print Lifter lifts higher quality impressions as the amount of dust residue transferred to the surface decreases. This presentation will give athe attendees new ideas and uses for the equipment they may have already or plan to purchase. This presentation will impact the forensic science community and/or humanity by allowing the forensic science community to understand what equipment should be used to obtain the highest quality footwear impressions from paper evidence. Crime Scene and forensic laboratory personnel are always trying to obtain the best evidence possible. This presentation * Presenting Author will give the forensic science community new ideas for developing footwear impressions on paper evidence and will give them a reference on which technique should be used. It will also show that if both techniques are used, what order they should be used in to obtain the best evidence. Therefore, this presentation will help the forensic community decide what techniques to use in order to obtain the highest quality footwear impression. The Electrostatic Dust Print Lifter is commonly used to lift footwear impressions from paper. The Electrostatic Detection Apparatus (ESDA), traditionally used to enhance indented writing, can also be used to develop footwear impressions on paper. This research compared the two methods for developing footwear impressions on paper in order to determine if both processes could be used to develop footwear impressions of the same or similar quality and in what order they should be used to develop the highest quality footwear impression. The sensitivity of each technique was also evaluated. The quality of the footwear impressions developed was determined by comparing twenty-five individual characteristics present on the known shoe to the footwear impressions developed using each technique. These footwear impressions were made by stepping on paper placed over several different surfaces. These surfaces included: linoleum, industrial Berber carpet, nylon carpet placed over a 3/8-inch pad, ceramic tile, cardboard, 1-inch foam, 4-inch foam, cement, asphalt, grass, and mulch. Each of the papers placed on these surfaces were developed using the Electrostatic Dust Print Lifter before the ESDA and using the ESDA before the Electrostatic Dust Print Lifter. The sensitivity test for the ESDA was conducted by placing ten sheets of paper (stacked) onto a carpeted hallway. This stack of paper was then stepped on with the known shoe. Each piece of paper in the stack, beginning with the top sheet, was processed with the ESDA until no footwear impressions were developed. The sensitivity test for the Electrostatic Dust Print Lifter was conducted by placing ten sheets of paper along a carpeted hallway. Each sheet was stepped on with the known shoe in succession beginning with the first sheet. Each of these sheets was processed with the Electrostatic Dust Print Lifter and compared. This study determined the footwear impressions developed using only the Electrostatic Dust Print Lifter were of better comparative value than impressions developed with only the ESDA. On average, 72.4% of the individual characteristics from the known impression were identified on images developed when only the Electrostatic Dust Print Lifter was used compared to an average of 38.9% when only the ESDA was used. Therefore, if only one technique is used, the Electrostatic Dust Print Lifter should be chosen. This study also determined if both methods are used on a piece of evidence, the ESDA should be used first and the Electrostatic Dust Print Lifter should be used second. This order results in satisfactory impressions with the ESDA and the Electrostatic Dust Print Lifter compared to using the Electrostatic Dust Print Lifter before the ESDA, which results in very low quality images with the ESDA. On average, 45.5% of the individual characteristics were identified using the ESDA first and the Electrostatic Dust Print Lifter second compared to an average of 72.4% using the Electrostatic Dust Print Lifter first and the ESDA second. Therefore, if the choice is available, the Electrostatic Dust Print lifter should be used instead of the ESDA. Unfortunately, if the Electrostatic Dust Print lifter is used first and the ESDA second, the results obtained using the ESDA will be of very low quality. The sensitivity test determined the ESDA develops high quality footwear impressions only on the top sheet of paper if the papers were stacked when the impression was placed on the paper. No footwear impressions were developed on any sheets under the top sheet of paper. The sensitivity test also determined the Electrostatic Dust Print Lifter results increase in quality as the amount of dust residue decreases on the surface. Therefore, crime scene technicians should be particularly interested in the top sheets of paper on a surface and footwear impressions in trace amounts of dust residue. These types of footwear impressions will most likely result in higher quality impressions that retain the most individual characteristics. Footwear Impression, Electrostatic Detection Apparatus, Electrostatic Dust Print Lifter 47 B33 Seasonal Variation of Phosphatase and Sulfatase Levels in Soil and Their Potential Use in the Comparison of Evidentiary Soil Samples Lawrence A. Quarino, PhD, and Janine M. Perna, BS*, Cedar Crest College, Forensic Science Program, 100 College Drive, Allentown, PA 18104 After attending this presentation, attendees will understand the possible utilization of enzyme activity in the comparison of soil samples. This presentation will impact the forensic community and/or humanity by helping with the development of a screening method for the comparison of soil samples. Enzyme activity in soil may vary according to geographical location and may be useful as a comparative tool in forensic geology. The type of location, weather conditions, and the season may influence the variability of enzyme activity. Thorton and McLaren (1) were the first to propose enzymatic characterization to differentiate soil. Their study, however, is somewhat limited since seasonal variation was not accounted for. This study is designed to examine the role of seasons and to better assess the use of enzymatic analysis for comparative purposes in soil. Soil samples were collected from four areas of differing topography and geography and assayed spectrophotometrically for the presence of two specific enzymes: phosphatase and sulfatase. Samples were taken from a location near water, an open field, a wooded area, and a sample from a hillside. Samples were collected during the spring, summer, fall, and winter. During each collection, five samples were taken fifty feet from a central point and mean values for enzyme activity determined. Statistical analysis using one way-analysis of variance at 95% confidence was conducted to determine significant differences in enzyme concentration among the sites. Greater variation exists between collection sites than within samples from the same site for each individual season. Results indicate that variation in enzyme concentration does exist between sites but the extent of the variation changes with season. Sample locations show greater differences in the spring and summer than in the fall or winter. The application of the method as a screening method used in conjunction with soil color comparison and particle size distribution will also be discussed. 1. Thorton JI, McLaren AD. Enzymatic characterization of soil evidence. J. of Forensic Sci. 1975; 20: 674-92. Soil, Phosphatase, Sulfatase B34 Detection and Identification of Personal Care Products in Sexual Assault Cases JoAnne Marzowski, PhD*, Washington State Patrol Crime Laboratory, 2203 Airport Way South, Suite 250, Seattle, WA 98134; Andrew W. Getz, Eastern Washington University, Cheney, WA 99004; and Kathy M. Geil, BS, MS, Washington State Patrol Crime Laboratory, 2203 Airport Way South, Suite 250, Seattle, WA 98134 The goal of this presentation is to detect and identify key ingredients of ointments, creams, lotions and personal lubricants on evidence submitted in sexual assault cases where there is no DNA present. This presentation will impact the forensic community and/or humanity by allowing the forensic scientist to determine if a personal care product was used in a sexual assault. This analysis may provide supportive case evidence and corroborate victim/suspect statements Personal care products such as ointments, creams, lotions, and personal lubricants used by assailants in sexual assault cases may serve as important evidence when there is no DNA present. Detection and 48 subsequent identification of key components of personal care products on clothing and in sexual assault kits may also provide supportive case evidence and corroborate victim/suspect statements. A representative sample of eighteen personal care products, including hydrophobic petrolatum based ointments, water based lotions, sunscreens, face and hand creams, were examined in this study. These products were smeared onto clothing and cotton swabs to simulate case evidence. A flowchart used for the detection of smears and analysis of key components of each type of personal product will be presented. This study describes the detection of smears on clothing and cotton swabs using a combination of visual observation, short and long wavelength ultraviolet light, the Forensic light source, and attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy. In addition, polarized light microscopy (PLM) is used to detect anisotropic smear components. This study also describes protocols for the extraction of smears from the substrates and the identification of key components using various analytical methods, including Fourier transform infrared spectroscopy (FTIR), gas chromatography / mass spectroscopy (GC/MS), pyrolysis gas chromatography / mass spectroscopy (pyrGC/MS), scanning electron microscopy /electron dispersive x-ray spectrometry (SEM/EDX), capillary electrophoresis, and/or high performance liquid chromatography (HPLC). Ointments Creams Lotions, Sexual Assault, FTIR GC/MS CE B35 Comparison of Pressurized Fluid Extraction Methods of Adsorbents For Use In Human Scent Evidence Collection Paola A. Prada*, Florida International University, University Park, CP345, Miami, FL 33199; and Alison M. Curran, BS, and Kenneth G. Furton, PhD, International Forensic Research Institute, Florida International University, University Park, Miami, FL 33199 After attending this presentation the attendee will learn the uses and applications of pressurized fluid extraction methods and their implementation for the purpose of the removal of organic volatile compounds from adsorbents used to collect human scent. This presentation will impact the forensic community and/or humanity by providing a better awareness for the selection of adsorbent materials used in the collection of human scent. The removal of the organic compounds present initially on collection materials will lead to a more “chromatographically clean” medium from which to analyze human scent for evidentiary purposes. The use of human scent identification by canines has been featured recently in a number of recent high profile court cases within the United States. While accepted in several European countries, the strict requirements for scientific evidence admissibility in the United States, set forth by the Frye and Daubert rulings have restricted the application of canine human scent identifications within the U.S. This is due in part to a lack of definitive studies demonstrating the reliability and validity of this approach, which has led to successful legal challenges to the use of these biological detectors in a court of law. As part of the scientific investigation into the forensic validity of canine identification of human scent, the chemical composition of scent must be proven to be unique to an individual and the scent profile must be proven to be stable over time. The instrumental determination of the organic compounds present in human odor plays an important role in determining the uniqueness of human scent, and thus the optimization of mediums for the collection of human odor is also of great interest. Presently, there are two main methods for the collection of human scent for the purpose of both instrumental and canine scent identification. The direct method is collecting the actual object, and the indirect method is collecting the odor on an absorber such as sterile gauze. Sterile gauze can be placed either in direct contact or placed close to the person or object of * Presenting Author interest. Air is also commonly drawn through the gauze pad in an attempt to increase the collection of odor. Canines have shown the natural ability to discriminate between odors in the presence of a high background, whereas instrumental analysis requires a significantly lower background. Despite sterilization through autoclave cleaning, the headspace analysis of sterile gauze pads using Solid Phase Microextraction with Gas Chromatography - Mass Spectrometry (SPME-GC-MS) has highlighted the presence of several organic compounds present within the sterile gauze prior to use. Thus sterile does not equate to chemically clean, and the contamination of the gauze observed has proven to be a serious limitation of the use of sterile gauze as an effective odor collection media for the instrumental identification of the compounds present in an individual odor profile. This paper discusses the use of carbon dioxide Supercritical Fluid Extraction (SFE) as a potential method of chemical cleaning of the gauze pads. A parallel study of the effect of SFE and Subcritical Water Extraction of absorbers will be presented. Optimization of the extraction parameters evaluated includes temperature, pressure and extraction time, and the use of chemical modifiers such as methanol and water will also be presented. Soxhlet extraction of absorbers through the use of various solvents, such as methylene chloride, methanol and chloroform, will also be presented. The chemical odor of gauze pads from various manufacturers and the absorbers currently used in European countries for the collection of scent evidence will be presented before and after SFE cleaning. This study is intended to demonstrate the importance of informed selection of gauze pads with minimal background odor, and those which can be cleaned most effectively to minimize any potential distracting odors from the gauze. With the use of chemically clean odor absorbers it is proposed that a better understanding of the composition of the chemical profile of human scent may be obtained, in turn benefiting the scientific acceptance of human scent identification by canine. Human Scent, Supercritical Fluid Extraction, SPME-GC/MS B36 Terminal and Secondary Effects of Bullets Fired Through Automobile Windshield Glass Ginger E. Eastham, BS*, Marshall University Forensic Science Program, 1401 Forensic Science Drive, Huntington, WV 25701; and Rodger R. Reed, BS, West Virginia State Police, 725 Jefferson Road, South Charleston, WV 25309 After attending this presentation, attendees will learn about data regarding trends in terminal ballistics and secondary effects of bullets fired through automobile windshield glass. These trends include effects on the glass, on witness panels, and in body armor vests that correlate to factors in bullet type, velocity, and weight retention. This presentation will impact the forensic community and/or humanity by demonstrating to forensic professionals and laymen how fired bullets react when passing through a glass obstruction, and the terminal results of said projectiles on the target. Law enforcemnt personnel will have a better understanding of ammunition characteristics in such a situation, and thus, will be better informed regarding choice of ammuniton to produce the effects desired by their agency. While many studies have been conducted to explore terminal ballistics, most have been in response to requests from military and law enforcement agencies for data concerning penetration and wound patterns. Additionally, several studies have been completed addressing the terminal ballistics of bullets fired into body armor. However, very little literature currently exists that seeks to observe effects of bullets fired through obstructions. In a laboratory environment, results obtained reflect ideal shots. In reality, many situations encountered by law enforcement personnel are much less than ideal. The high stress nature of such moments has a high * Presenting Author impact on officers and suspects alike, and can cause widely varying results in reaction time and aim. The environs of the conflict further compound altercations of this nature. In other words, most situations occur as parties are firing at one another from behind some solid object or glass rather than a one-on-one confrontation on open ground. Any obstacle in the bullet path will change the effects of that bullet upon its final target, if it reaches the target at all. A controlled environment is necessary to observe any phenomena or trends of bullets fired through solid objects, but this will necessarily reduce the random nature of the very act the experiment is trying to replicate. In particular, this experimentation was geared toward recording effects produced by firing through a glass obstruction. Glass was deemed a better material in which to visualize effects while keeping incidence of low angle impact (ricochet) low. Such cases involving glass are often experienced in shootouts as well as on the highway during driveby shootings. One such scenario encountered by law enforcement personnel occurs when they use their vehicles for cover from fire. With the support of the West Virginia State Police and its Firearm/Tool Mark section laboratory, this scenario was replicated for study. Shots were fired from approximately six feet away through a windshield and into a stabilized body armor vest. The setup was accomplished in part from the generous donation of windshields from a local supplier. This poster will display the examined effects of bullets fired through an auto glass obstruction as well as the affect of velocity and weight retention upon the observed results. Additional factors in ammunition performance such as bullet type and manufacturing process are discussed. Seventeen forty-caliber (.40) common-duty ammunitions currently in use by law enforcement agencies were fired through glass windshields and witness panels into body armor. Velocity and weight retention as well as diameter of all bullet holes and other secondary effects were measured and recorded. Control tests of body armor vests as well as bullet recovery tank testing were also performed. Study of multiple test fires revealed general trends in bullet interaction with the windshield glass, the witness panel, and the body armor, respectively. Type of bullet played a significant role in the presence or lack of fragmentation. Of all brands tested, Federal Cartridge Co. Tactical law enforcement ammunition produced the highest area and approximate volume of damage upon impact with target. However, the philosophy of a particular agency or a particular situation in which shots are fired may determine which ammunition should be used to obtain desired result. Terminal Ballistics, Firearms, Glass B37 LA ICPMS and IRMS Isotopic and Other Investigations in Relation to a Safe Burglary Gerard J.Q. van der Peijl, PhD*, Netherlands Forensic Institute of the Netherlands, Ministry of Justice, PO Box 24044, The Hague, 2490 AA, Netherlands; and Shirly Montero, PhD, Wim Wiarda, Ing, and Peter de Joode, Ing, Netherlands Forensic Institute, PO Box 3110, Rijswijk, 2280 GC, Netherlands After attending this presentation, attendees will appreciate the strong discriminating power of isotopic technique such as (LA) ICPMS and IRMS, not only for these specific safe burglary investigations but for a wide spectrum of other forensic investigations. This presentation will impact the forensic community and/or humanity by making forensic community further aware of the strong power of LA ICPMS and IRMS isotopic techniques. Safe wall filling material was released when an old safe was burglarized. During a search at the suspect’s residence, visually similar material (mixture of glass like particles and sawdust) was retrieved from a plastic bag containing money. Small iron-containing particles were also retrieved from both samples. The safe wall filling material was found to 49 consist of a mixture of alum or potassium alum (KAl(SO4)2.12H2O) crystals (XRF/XRD) and sawdust (vis, FT-IR). All three material fractions (alum, sawdust, metal particles) were intercompared for the two samples. The material combination of alum and sawdust appears very rare. Information was collected on the history of its use as safe wall filling, alternative applications of this material combination and how often Dutch police organisations encountered this material in the last two years. It was concluded that this material combination was only used for safe wall filling in the period before the 2nd World War, that no alternative applications were found and that no police organisation reported encountering this material combination. Alum investigations: The glass like particles in both material samples were classified as alum. ICP AES results were obtained using a PE OPTIMA 3000 instrument. Apart from the major elements Al and K (S was not measured) Cr and Tl were observed at similar levels in both samples. Other elements observed (Fe, Sr,..) were present at much higher concentrations in the red-brown powder so that these are not confidently attributed to the alum samples. LA ICPMS results using a PE ELAN 6100 DRC PLUS instrument were obtained on freshly cleaved inner alum crystal surfaces. Elements observed in both samples are Mg, Si, P, Ti, Cr, Cu, Zn, Ga, Rb, Sr, Sn, Ba, Tl and Pb. IRMS services were provided by Iso-Analytical Ltd (Sandbach, UK) using standard methods on a Europa Scientific Geo 20-20 instrument with an EA-IRMS interface. Different isotope ratios were measured in separate experiments using different experimental configurations of the instrumentation. The mean δ34S values for the suspect and crime scene alums (both 2.2 ± 0.1 ‰, n = 6) were undistinguishable using an experimental uncertainty of 0.24 ‰ (2s). This is compared to a variation range of -3.9 to +30.6 ‰ as reported [1] for δ34S values obtained for a large number of alum minerals from various locations throughout the world and a variation range of 1.7 to 9.8 ‰ for a small test set of four different alum samples as obtained from alum suppliers in the Netherlands. Sawdust investigations: The suspect and crime scene sawdust samples are visually similar and exhibit the same red brown color. A mixture of wood species, both soft and hard woods, was observed in both samples during botanical species investigations at the National Herbarium of the Netherlands. Over five species were identified in the suspect sample. IRMS isotope ratios δ2H, δ13C en δ18O for both the suspect and the crime scene sawdust fractions agree within the experimental uncertainty (2s, n=6). δ13C=-25,71+ 0,15 is e.g. compared to a variation range of -32 to -22 ‰ as reported for the δ13C values of C3 plants [2]. Applicability of this information for the local situation is tested by collecting a limited set of sawdust samples in the Netherlands market and analysing these by IRMS. For δ13C e.g. a variation range of -23 to -28 ‰ is measured. Metal particles investigations: In both samples many minute metal particles were observed that were magnetically separated from the sample matrix. Corresponding variations in morphology of the particles (round balls of various sizes and curved lint like particles) indicate that metal particles in the two samples were formed through a similar process consistent with abrasive cutting of a safe [3]. µ-XRF (EAGLE) results for the round balls and lint particles show a high abundance of Fe as well as the presence of other elements such as Al, Si, P, S, Mn and Cu. Similar results were obtained for the fine metal particle fractions in both samples. Conclusions: Combining the information from the various sources and the analytical results for the various material fractions (sawdust, alum, metal particles) provides a strongly discriminating method to compare the (combination of) material fractions as found in the two samples at the suspect’s residence and at the scene of crime. References 1. Rye, R.O., P.M. Bethke and M.D. Wasserman, The Stable Isotope Geochemistry of Acid Sulphate Alteration, Economic Geology 87 (1992) 225-262 2. Cerling, T. E., J. M. Harris, B. J. MacFadden, M. G. Leakey, J. Quade, V. Eisenmann and J. R. Ehleringer, Global change through the Miocene/Pliocene boundary. Nature 389 (1997) 153-158. 50 3. Zeichner A., G. Feingold and E. Landau, “Abrasive cutting of a safe: a case study”, Journal of Forensic Science 38 (1993) 1516-1522 ICPMS, IRMS, Safe B38 An Analytical Approach to Comparative Bullet Lead Analysis: Physical and Chemical Aspects of Discrimination Diana M. Wright, PhD*, Charles A. Peters, BS, and Marc A. LeBeau, MS, FBI Laboratory, Chemistry Unit, Room 4220, 2501 Investigation Parkway, Quantico, VA 22135 After attending this presentation, attendees will understant the methodology used for the physical and chemical analysis of bullet lead specimens. The protocol, method validation, report terminology, and significance assessments generally conveyed in testimony regarding positive associations of comparative bullet lead evidence will be presented. This presentation will impact the forensic community and/or humanity by educating the forensic community to the steps taken to validate a protocol for quantitative analysis of a manufactured product. It will describe how the protocol was validated and provide the community with examples of the language used in reports and during testimony to convey the conclusions drawn from the analysis. Comparative bullet lead analysis is the physical and chemical examination of lead bullets, fragments, or shot pellets. It is performed when a firearm is not recovered or if a bullet or lead fragment lacks sufficient marks of value for a firearms examiner to determine conclusively that the specimen can be associated with a specific firearm. This examination is not as probative as a positive association between a firearm and the ammunition it discharged. However, comparison of recovered cartridges to the bullets or fragments from the victim or scene can provide a circumstantial link based on the ammunition used in the commission of a crime. This poster will describe the most current protocol used by the FBI Laboratory to perform this examination, which has been used in support of local and federal law enforcement investigations for over 35 years. While the sample described in this work requires certain parameters, the methodology can be easily adapted to analysis of any of the wide range of specimens increasingly encountered in modern forensic laboratories in the 21st century. For example, small modifications would allow the digestion procedure to be readily adapted to the analysis of glass or steel specimens. The protocol begins with a physical comparison of the items submitted. Many discriminating features are commonly found on ammunition that can be used to disassociate items quickly and conclusively. These differences and the documentation necessary to describe them will be discussed. In the absence of physical discriminators, a chemical analysis of the submitted lead specimens is required. The procedure will be described in detail to include: documentation of the physical alterations necessary to sample the evidence; the chemical digestion process; quality assurance and controls; and instrumentation used. Validation data for the chemical analysis of bullet lead specimens will be offered for consideration. The parameters that were measured in validating this method include: limits of detection and quantitation, accuracy, within-run precision, between-run precision, precision using separate analysts for data collection, linear range, sensitivity, and analyte recovery. Results will be offered for a sample case in which the physical and chemical examinations could not differentiate between two or more specimens. The language used to report these findings will be provided. Lastly, the conclusions or interpretation that a qualified examiner would place on these findings will be described as a guideline for the strength and limitations of quantitative analysis applications to commercially - available manufactured products. Bullet Lead Analysis, Compositional Analysis, Comparative Examinations * Presenting Author B39 Forensic Elemental Analysis of Glass by Laser Induced Breakdown Spectroscopy (LIBS), Laser Ablation ICPMS, and X-Ray Fluorescence Sayuri Umpierrez, BS*, Waleska Castro, MS, and Marilyn Prieto, BA, Florida International University, Department of Chemistry and Biochemistry and International Forensic Research Institute, 11200 Southwest 8th Street CP 194, Miami, FL 33199; Scott Ryland, BS, Florida Department of Law Enforcement, 500 West Robinson Street, Orlando, FL 32801; and José R. Almirall, PhD, Florida International University, International Forensic Research Institute, Department of Chemistry and Biochemistry, 11200 Southwest 8th Street, CP 316, Miami, FL 33199 After attending this presentation, attendees will learn about different solid sampling techniques for elemental analysis: laser induced breakdown spectroscopy (LIBS), laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), scanning electron microscopy with an energy dispersive spectrometer (SEM EDX) and x-ray fluorescence (XRF). The advantages, disadvantages, and application of these techniques to forensic casework will be discussed. This presentation will impact the forensic community and/or humanity by demonstrating the different solid sampling techniques for elemental analysis: laser induced breakdown spectroscopy (LIBS), laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), scanning electron microscopy with an energy dispersive spectrometer (SEM EDX) and x-ray fluorescence (XRF). Materials analysis and characterization can provide important information as evidence in legal proceedings. Although the utility of trace elemental analyses for comparisons of glass, paint fragments, bullet lead and metal fragments has been shown to offer a high degree of discrimination between different sources of these materials, the instrumentation required for the generation of good analytical data in forensic comparisons can be costly and require a high degree of analytical sophistication. Refractive Index (RI) has been used for glass comparisons in combination with elemental analysis using a variety of methods of solid sampling, including; Scanning Electron Microscopy with an Energy Dispersive Spectrometer (SEM-EDX), x-Ray Fluorescence (XRF), Laser Ablation Inductively Coupled Plasma Atomic Emission Spectroscopy (LA-ICP-AES) and, more recently, LA-Inductively Coupled Plasma Mass Spectrometry (LA-ICPMS). A newly developed Laser Induced Breakdown Spectroscopy (LIBS) instrument (Foster and Freeman Ltd., Evesham, U.K.) has been evaluated as a tool for the forensic elemental analysis of glass and compared in performance to other elemental methods in order to determine the utility of comparing casework sized glass samples. Developments in instrumental design of the LIBS system are presented. The discrimination power afforded by the elemental analysis of a sample set having similar refractive indices measured by the LIBS system is reported in contrast to that provided by micro-XRF and LA-ICPMS. The advantages and disadvantages of using these solid sampling elemental analysis techniques will also be presented. Solid Sampling Elemental Analysis, Glass, LIBS B40 Choosing a Statistical Method for the Data Assessment of the Compositional Analysis of Bullet Lead Diana M. Wright, PhD*, FBI Laboratory, Chemistry Unit, Room 4220, 2501 Investigation Parkway, Quantico, VA 22135; Robert D. Koons, PhD, JoAnn Buscaglia, PhD, and Heather L. Peters, PhD, FBI Laboratory, Counterterrorism and Forensic Science Research Unit, FBI Academy Complex, Quantico, VA 22135; and Marc A. LeBeau, MS, FBI Laboratory, Chemistry Unit, Room 4220, 2501 Investigation Parkway, Quantico, VA 22135 The goal of this presentation is to provide a description of five statistical approaches to trace element concentration data is provided. These methods were compared for 93 randomly selected cases. Results will describe how each approach grouped the case data and explain which approach was ultimately selected for use in current casework. This presentation will impact the forensic community and/or humanity by serving the forensic community through the discussion of several statisitical approaches that one may utilize in determining acceptable match criteria for quantitative analysis of man-made products. The FBI Laboratory is one of the few forensic laboratories that perform quantitative trace element analyses on a routine basis for a wide variety of specimens, including glass, steel, and bullet lead evidence. The bullet lead protocol was specifically developed by the FBI Laboratory over 35 years ago to assist local and federal law enforcement agencies with shooting investigations where a firearm is not recovered or the fired bullets are badly damaged. The protocol, including the association criteria used to determine if two or more specimens are analytically indistinguishable, has evolved over the course of this examination’s history. As the 21st century progresses, the FBI Laboratory has revised its analytical protocol for comparative bullet lead analysis. Part of this revision involved a review of the statistical approach used to determine associations. To that end, this poster will present work that has recently been completed through the collaborative efforts of the FBI Laboratory Division’s Chemistry Unit and the analytical research group of the Counterterrorism and Forensic Science Research Unit. Five statistical approaches were applied to the quantitative results collected for 93 randomly selected comparative bullet lead cases that have been submitted to the FBI Laboratory over the last decade. The purpose of this study was to determine if any of the results reported out to contributors using the established criterion would have been affected if a different statistical approach had been used. The bullet lead protocol for the past decade has employed an association criterion that requires the measured precision of the specimens being compared to overlap within the range of 2. Therefore, in order for two specimens to be considered analytically indistinguishable, an overall range of 4 was applied to the concentration results. The five methods compared in this study were: range overlap, 2, the student t test, the successive t test, and Hotelling’s T2 test. The first three methods utilized measured precisions associated with the specific case results as a basis for establishing match criteria. The latter two statistical assessments required the use of pooled historical data in order to calculate precision measurements for the necessary equations. Any assumptions that were made in order to perform the testing on the pooled results will be described. Results will be presented for the five tested analytical approaches as well as the conclusions issued in the corresponding reports. Ultimately, there were no cases in which a Q/K association was reported in the absence of corroborating associations from at least one of the tested methods. The method chosen for the revised protocol will be described and the rationale for its use will be addressed. Statistical Analysis, Bullet Lead Analysis, Comparative Examinations * Presenting Author 51 B41 Evaluation of Solvent Systems and Mobile Phases for the Extraction and Identification of Fiber Dyes by Liquid Chromatography Mass Spectrometry (LC-MS) Michael E. Sigman, PhD*, Min Huang, PhD, and Breanne Hornsby, National Center for Forensic Science, University of Central Florida, PO Box 162367, Orlando, FL 32816 The objective of this presentation is to demonstrate the advantages of liquid chromatography – mass spectrometry (LC-MS) in the comparison of questioned and known fibers. The compatibility of LC-MS methods with existing dye extraction protocols will be emphasized. This presentation will impact the forensic community and/or humanity by emphasizing the need for enhanced methods for the analysis of fiber evidence and demonstrating the compatibility of LC-MS methods with existing FBI SWGMAT dye extraction protocols. Previous research has shown that cotton fibers that are indistinguishable by means of microspectrophotometry, HPLC analysis and FBI SWGMAT dye extraction protocols, may be discriminated by LC-MS analysis of the extracted dyes. Mass spectrometry provides a molecularlevel interrogation of the dye structure and, as a result, allows the differentiation of dyes having differing molecular structures. The same level of discrimination is not possible based only on UV-visible absorption profile, chromatographic behavior or extraction characteristics. The previously reported LC-MS analysis methods have been extended by evaluating their compatability with the solvent systems recommended by FBI SWGMAT protocols for fiber dye extraction. Solvent systemts that were evaluated for LC-MS analysis included pyridine/water, formic acid/water, acetic acid, chlorobenzene, chloroform, and dimethylformamide (DMF)/formic acid (1:1). The dyes that were extracted in each solvent system were readily chromatographed in a methanol/water gradient with the extraction solvent exerting no significant influence on the chromatographic behavior of the dyes. The chromatographic behavior of the dyes was found to be significantly influenced by the addition of 0.1% acetic acid or trifluoroacetic acid to the mobile phase for the analysis of basic dyes in positive ion mode. Similar enhancements were obtained by the addition of TEA (triethylamine) to the mobile phase for the analysis of acidic dyes in the negative ion mode. In both cases the additive enhanced sensitivity and separation efficiency of the dye by ESI-MS analysis and reversed phase chromatographic separation. The analyses were conducted on standards comprised of dyes of known structure that had been commercially applied to fibers of known composition. This presentation will emphasize the need for enhanced methods for the analysis of fiber evidence and demonstrate the compatibility of LC-MS methods with existing FBI SWGMAT dye extraction protocols. Fiber Analysis, Trace Evidence, Mass Spectrometry B42 The Effects of Environmental and Atmospheric Conditions on the Longevity of Latent Prints Michele Eichenmiller, BA*, Marshall University-Forensic Science Center, 1401 Forensic Science Drive, Huntington, WV 25701; Steven King, BA, West Virginia State Police-Latent Print Section, 725 Jefferson Road, South Charleston, WV 25309; and Pamela Staton, PhD, Marshall University-Forensic Science Center, 1401 Forensic Science Drive, Huntington, WV 25701 After attending this presentation, attendees will learn about the effects of weather and humidity on the permanence of latent prints. 52 This presentation will impact the forensic community and/or humanity by providing insight into the effects of different atmospheric and environmental conditions on latent prints as they are exposed to varying environments over a specified time. This scientifically gained information can then be referenced in court proceedings when latent print examiners are called to testify about the longevity of latent prints. As latent fingerprints are deposited on a surface, several factors may affect their recovery particularly as they age. Such factors include subject, substrate and matrix, and environmental factors. “Subject” factors include age, sex, stimuli, occupation, and medical conditions. For example, a car mechanic with motor grease on his hands would leave a greasy latent print that would resist degradation better than a print made up of mostly sweat, as left by a person who has been exercising. In general, latent prints are composed of 99% water and 1% oil secretions. Some factors can be accounted for by the latent print examiner but other influences affecting the deposited print are unknown. The examiner does not generally know subject factors, such as age and medical conditions, which may influence the hardiness of the deposited print. “Substrate” factors are associated with the distortion or interference created by the surface on which a print is deposited. This is demonstrated by textured surfaces’ inability to retain a good impression as opposed to a smooth surface, which would retain a good latent print. Matrix factors are those involving the material making up the friction ridge skin impression and can be deduced and assessed to some point from the submitted evidence. An example of a matrix is the motor grease in the previous example. While potentially important, examiners rarely receive information associated with atmospheric and environmental factors that may impact the latent print at the crime scene or on the evidence as it ages. These variables include temperature extremes, humidity, and significant weather events such as rain that may have influenced the permanence of the latent print. All of these variables can interact with the water and oil content of the latent print. This interaction will thereby affect how well the print is preserved and how quickly it will degrade before it is rendered completely useless for comparison purposes. The effect of atmospheric and environmental factors on the permanence of latent fingerprints is an often-raised question that latent print examiners must face as they testify in court proceedings. Up to this point, examiners have relied on personal knowledge to convey to the court their opinion with respect to the longevity of fingerprints. This experience is not based on peer-reviewed scientific research but rather on the experience of the examiner. In fact, when a literature search is conducted of current journal articles, few studies are identified which document the effects of different environmental and atmospheric factors on a variety of substrates and matrices or on the life of a latent print left on such surfaces. This study shows the effects of an indoor versus outdoor environment on latent prints over the course of four weeks. Latent prints were deposited on a variety of porous and nonporous surfaces and then placed either inside the building or outside, in a sheltered location. Four types of prints were left on each surface and then collected once a week for four weeks. The four types of prints were clean, natural oils, artificial oils, and dirty prints. The results of this experiment show a pattern of marginal latent print longevity on porous surfaces exposed to summer heat and humidity. Prints on nonporous surfaces also exhibited decreased longevity when exposed to outdoor conditions. The permanence of latent prints decreases as a result of the time spent outdoors. This is in opposition to the same surfaces kept in an indoor environment at a constant temperature and humidity, which exhibit greater permanence in regard to the latent prints, regardless of time spent indoors. This poster will present the work conducted at the West Virginia State Police Forensic Laboratory-Latent Print section during a summer internship. This internship is to fulfill the requirements of the Marshall University Forensic Science Program Summer Internship. Latent Prints, Longevity, Environmental Conditions * Presenting Author B43 Volatile Compounds Produced by Decomposing Human Blood and Those Detected by Cadaver Dogs This study proposes that the volatile chemicals detected during the decomposition of human remains must also be present during the decomposition of human blood. The use of fresh blood that is permitted to decompose naturally, versus expired blood or plasma from a blood bank, has the potential to affect the decomposition odor observed. Wilma Jeffris, BA*, Samantha Tolliver, BS, and Kenneth G. Furton, PhD, Florida International University, University Park Campus, CP 345, Miami, FL 33199 SPME-GC-MS, Blood Decomposition, Cadaver Dogs After attending this presentation, attendees will earn about the volatile organic compounds that are produced as human blood decomposes and the odor signature compounds cadaver dogs alert to. By determining the odor signature compounds produced in aging blood, better and less hazardous training tools can be developed. Better training will lead to more reliable detection of human remains and trace blood spatters not readily visible and identification of crime scenes where bodies are not present. This presentation will impact the forensic community and/or humanity by assisting in the identification of the odor signature compounds which will lead to better trained cadaver dogs, more reliable evidence gathered, better results in finding bodies, and future development of instrumental methods with equal or better detection than detector dog teams. Cadaver detection canines are dogs trained specifically to locate human remains. Although the use of canines as detection methods for items of forensic interest has been accepted historically for many years, the Frye and Daubert rulings of forensic evidence admissibility require validation of scientific processes. Accordingly, the canines must be capable of locating these remains in many environments, but must not be confused by the decomposition of other animal species. Therefore, the training protocols of most law enforcement agencies stipulate the use of human remains and human blood, both in various degrees of decomposition. Although these training practices are generally accepted within the canine handling community, the safety concerns regarding the biohazards of human tissue, and the possibility of infection of blood-borne disease such as HIV/AIDS, and hepatitis B/C confirms that the training aids used by handlers do pose a significant health risk. Currently available pseudoodors, those that mimic human decomposition, have been shown to be of limited application to canine training. Training aids that effectively reproduce the odor of decomposition without the associated risk of blood handling have the potential to replace real tissue aids in most circumstances, while maintaining the high standard of odor discrimination. The process of blood decomposition involves coagulation of the blood, and thus may be limited or modified by the addition of anticoagulants such as EDTA and CPDA1 to the blood samples to extend shelf life or prevent decomposition during storage. To date, the role that each component of blood plays in the decomposition process has not been determined, nor have the compounds formed been identified. However with coagulation playing an important role in the overall process, it is proposed that the plasma component of blood is heavily involved. Samples of blood and blood components, both with and without anticoagulants, have been allowed to decompose over time. The headspace of these samples has been collected and analyzed using Solid Phase Microextraction (SPME) with Gas Chromatography - Mass Spectrometry (GC-MS). Blood itself is a complex mixture of plasma and cells, hence given the complexity of the blood sample, and the diverse nature of the compounds present, polydimethylsiloxane (PDMS) and polydimethylsiloxane/divinylbenzene (PDMS/DVB) SPME fiber chemistries are combined in complementary analysis. Several variables were considered during the development of a technique that will provide an efficient means of identifying the volatile compounds. Blood and blood component samples were kept in various temperatures including room temperature, 37 degree Celsius, and under excessive heat. Seven-year-old, one-year-old, three-month-old, and fresh blood samples were analyzed to determine the development and disappearance of the desired volatile compounds in the presence and in the absence of anticoagulant. B44 * Presenting Author Examination of Counterfeit Hong Kong $10 Bimetallic Coins Nai-Chiu Kwok, BSc, MPhil*, Fu-Chiu Kwok, PhD, and Koon-Hung Wong, PhD, Goverment Laboratory, 8/F, 88 Chung Hau Street, Homantin Government Offices, Hong Kong, Nil Nil, China After attending this presentation, attendees will learn a general guideline of examination involving the physical parameters and surface features in examination of bimetallic coins. This presentation will impact the forensic community and/or humanity by providing an examination guide on bimetallic coins including the core-extrusion method which is an effective and reliable supplementary method of examination. Moreover, the core-extrusion method may also be applied to coins with similar construction. This poster presents a number of important physical parameters, such as surface features of the genuine bimetallic HK$10 coins which can be utilized for the examination of counterfeit coins. The Hong Kong ten-dollar coin was introduced into common circulation by the Hong Kong Monetary Authority towards the end of 1994. However, just within six month after launching, counterfeit HK$10 coins started to be found in the region. The first case of counterfeit HK$10 coins was delivered to the Hong Kong Government Laboratory in the summer of 1995. Since then, there were significant increases in the number of cases and the actual number of coins submitted for examination. The quality of counterfeit ten-dollar coins which were first found was crude. They displayed significant difference, both in physical dimension and/or surface features, to the control specimen. However, as time progressed, forgery techniques appeared to have improved to such an extent that counterfeit HK$10 coins were almost indistinguishable to the untrained eye from the corresponding control coins. General guidelines for the examination of HK$10 coins are as follows: 1) The physical parameters of the coin, such as weight and dimensions (thickness and/or overall diameter) are measured, but as the physical parameters of counterfeit coins can most easily be reproduced within a reasonable level of accuracy, thus they would not provide a sound indication of authenticity. 2) The surface and edge characteristics of the counterfeit coin are scrutinized and compared with genuine coins. For example, the color of the bimetallic part is distinctly different from the control specimen, the bonding between the core and the annulus is rough usually; the inscribed words are of different fonts; the milling is poor; the Bauhinia logo lacks clarity and details. In most of the cases, visual examination would normally be sufficient to distinguish a counterfeit coin from a genuine control. 3) Determination of the metal composition of the coins can also be used. Accurate results can be obtained using such techniques as Atomic Absorption Spectroscopy (AAS) or Inductively-coupled Plasma-Atomic Emission Spectrometry (ICP-AES). However, these techniques are destructive, labour-intensive and timeconsuming as they involve pre-treatment of sample. An alternative method being used is the Scanning Electron Microscope (SEM) – a non-destructive, reasonably accurate technique that requires minimal sample treatment. Recently, an effective physical method (core-extrusion method) has been developed in the authors’ Laboratory. It makes use of the construction 53 of the coins itself. The HK$10 coin is bi-metallic consisting of a core made of nickel-brass with a cupro-nickel annulus. The two metallic parts are held together by a physical construction consisting of a groove at the outerperimeter of the core and a rim at the inner perimeter of the annulus. Therefore, for the genuine coin, a greater force exerted by way of a hydraulic press on the core will be required to separate it from the annulus rim. By comparison that required for separating the core and the rim of a counterfeit coin is only 20% of that for the genuine counterpart. By far the most time-consuming step in the analysis of HK$10 coins is the metal-composition determination. Whereas the most effective way to distinguish a counterfeit HK$10 coin would be visual examination of the surface features, such as the Bauhinia and the word fonts. Moreover, the core-extrusion method also serves as an effective and reliable supplementary method of examination. The latter method may also be applied to coins with similar construction. Counterfeit, Bimetallic, Coins B45 An Evaluation of the Trigonometric Model for Point of Origin Prediction in Bloodspatter Pattern Analysis Anna S. Duggar, MS*, and Rose R. Duryea, MA, New Orleans Police Department Crime Laboratory, 2932 Tulane Avenue, New Orleans, LA 70005 After attending this presentation, attendees will learn the responsible use of the trigonometric model in applied fieldwork and the value of training by a qualified expert. The discipline of bloodspatter pattern analysis continues to need peerreviewed scientific and statistical studies, particularly on essential methods. This presentation will impact the forensic community and/or humanity by meeting that need and providing quantitative arguments for training and responsible method implementation. The discipline of bloodspatter pattern analysis continues to need peerreviewed scientific and statistical studies, even on premises as essential as the trigonometric formula historically used in the back-calculation of a point of origin from individual droplet stains in an impact pattern. Structured, objective scientific research in this field in the last two decades has shown that even the most basic assumptions are subject to change as understanding of the science is explored. In addition, recent judicial challenges to the testimony of the bloodspatter analysis expert as neither scientifically valid nor admissible, combined with Daubert challenges to other pattern analysis fields of the forensic sciences, have produced a demand for statistical support for the conclusions of the expert analyst. The experiment presented here describes the confidence interval and relative accuracy of the trigonometric calculation used in point-of-origin reconstruction for bloodspatter patterns. To best test the mathematic model, it was necessary to produce a large number of stains at a known, constant angle. To this end, an apparatus was built allowing for controlled replication of single droplet patterns in bovine blood at a range of incident angles between zero and 80 degrees. The stains were then measured by multiple researchers, both trained and untrained, and the length and width measurements compared. These measurements were used to calculate the angle of incidence using the formula ∠incidence = cos -1 (d / D). The error of the calculated angle as compared to the recorded angle was determined through scatterplots and other statistical analysis. Results support the conventional wisdom that as the ratio of width to length of the stain approaches 1.0 (impact angle approaches 90 degrees), calculations become less reliable. The impact of even minimal training by a qualified expert is demonstrated, and comparison of the range of calculated angles to the ideal calculated angle provides new insight into the responsible use of the trigonometric model in the field. Bloodspatter, Bloodstains, Statistics 54 B46 The Accreditation of College and University Forensic Science Programs Jay A. Siegel, PhD*, Indiana University, Purdue University, School of Science, LD 326, 402 North Blackford Street, Indianapolis, IN 46202 After attending this presentation, attendees will learn how the process of accreditation is carried out. This presentation will impact the forensic community and/or humanity by demonstrating how the quality of forensic science education will be improved. The American Academy of Forensic Sciences in conjunction with the National Institute of Justice has begun a process for accrediting College and University Forensic Science Programs by adopting a set of guidelines developed by the Technical Working Group on Forensic Science Education. This poster will show the process and procedures for accreditation. Forensic Science, Education, Accreditation B47 Proof of a Negative? H. Dale Nute, PhD*, Florida State University, 4750 Collegiate Drive, Panama City, FL 32405 After attending this presentation, attendees will learn an approach to teaching the concept of negative evidence to students or trainees using crime scene search protocols as exemplars. This presentation will impact the forensic community and/or humanity by introducing forensic science practitioners and educators to the concept of constructing a probability assessment for a crime scene search protocol, in particular to the probability that a negative search can, under the right circumstances, infer a probability that the evidence was not there. Determining the error rate for many forensic science examinations is relatively straightforward. For example, the probability of correctly: identifying a compound, assigning a DNA pattern to its donor, and reconstructing a vehicle’s speed at the time of an accident, have all been considered. But, what if a crime scene search produces no evidence? What is the probability that it really was there (merely overlooked) versus the probability that it really was not there? What is the approach for calculating the error rate for a crime scene search such that one can offer a reliable assertion that an item of interest really was not there when its absence will support an issue in the case, i.e., negative evidence? The reliability associated with locating, or assuring the absence of, an item of evidence is no less trivial than the reliability associated with its examination. The question then is, what is the probability that a particular search protocol will locate (or miss) a particular item in a particular matrix at a scene? This question is analogous to the concept of limit of detection in toxicological analyses. And, like those tests, the answer depends on the compound of interest, the matrix within which it is hidden, and the technique used to detect it. But, unlike the relatively uniform approaches associated with identifying chemical compounds in body fluids, the variety in forms of evidence, types of matrices, and detection techniques that comprise crime scene searches precludes simple solutions. Indeed, like much of forensic science, the question can sometimes be answered quantitatively but more often it can only be answered qualitatively. Faced with all of this complexity, the educational challenge is first to convey the concept itself. An approach is presented for teaching students how to consider the question of presence versus absence. An exercise is constructed around the task of locating small items, such as cartridge cases, in an outdoor plot. A variety of search techniques is employed with the varying successes recorded. From these data, a comparison can be constructed as to the relative probabilities of finding the item of interest. One also can determine the relative “cost” in terms of time and equipment required. These findings * Presenting Author lead to another question – What would be the increase in the probability of finding the item(s) if two different techniques were employed instead of just one? This question allows the instructor to introduce the idea of Bayesian probability as applied to an examination, rather than just to its impact on the case as is commonly presented. Considering a Bayesian approach, though, requires a consideration of both false positives and false negatives along with the usual consideration of true negatives and positives. Using cartridge cases as an exemplar allows for plausible scenarios to be advanced in which a probability exists of finding items that have nothing to do with the case at hand. This introduces the idea of combining quantitative data from objective sources with “expert,” “background,” estimated, thus, subjective information. Although the students do not have the skills, based on the exercise alone, to develop reliable data as to a search protocol’s error rate, they know the procedure and have a better understanding of what it takes to “prove a negative.” Negative Evidence, Crime Scene Search, Protocol Error Rate B48 Unusual Use of Modified Fire Debris GC/MS Method to Analyze Hydrocarbons Elzbieta J. Kubicz, PhD*, Wyoming State Crime Laboratory, 316 West 22 Street, Cheyenne, WY 82002 After attending this presentation, attendees will learn the adjustment of known GC/MS analysis of fire debries to identify common hydrocarbons. This poster will present the unusual use of GC/MS method for analysis of fire debris. This presentation will impact the forensic community and/or humanity by demonstrating easy modification of existing method to analyse hydrocarbons. Case History: The 4-year-old male was found unresponsive inside a tent in the yard of his home. The 911 call was placed about 1:00 p.m. A temperature reading was taken inside the tent, and registered 126°F. The child was taken to egmergency room. The rectal temperature at that time was 108°F. The child was pronounced dead at 1:39 p.m. Toxicologic evaluation identified Ethylbenzene and Xylene in a blood at 0.85 mcg/ml and 4.1 mcg/ml respectively. Cause of death was ruled acute intoxication. The boy’s parents claimed that they were removing lice using “Ortho” and “Hot Shoot” sprays. To preserve the evidence, immediate analysis was needed. The child’s bedding from the tent was subject to analysis to look for trace of Xylene and Ethylbenzene. Ethylbenzene, GC/MS, Xylene B49 Reliability Testing of Commercial Containers for Fire Debris Evidence Storage Michael E. Sigman, PhD*, Mary R. Williams, BS, and Denise Fernandes, BS, University of Central Florida, PO Box 162367, Orlando, FL 32617 The goal of this presentation is to present results from a study on the leak rates of ignitable liquids from commercial containers used for fire debris evidence storage. This presentation will provide information on the reliability of commercial containers as fire debris evidence containers and methods for handling and analyzing the evidence. * Presenting Author This presentation will impact the forensic community and/or humanity by presenting the results from this long-term study by monitoring hydrocarbon retention in quart volume containers of metal cans, glass jars, and polymer bags. Data revealing leak rates of the containers and the behavior of the hydrocarbon vapors will be discussed along with the significance to methods in fire debris collection and analysis. A common type of evidence collected at a fire scene is the debris suspected of containing ignitable liquids. The total or partial loss of the volatile ignitable liquids in a container subject to leaking can lead to an altered hydrocarbon profile. Previous investigations have reported significant leak rates for various commercial containers primarily paint cans and polymer bags. This study will differ from previous studies based on its approach for determining the leak rates of the commercial containers for fire debris evidence. One approach utilized in this study was based on the ideal behavior of a hydrocarbon solution in the C7-C10 range in accord with Raoult’s law. Leaks in the container would result in a change in vapor phase composition and a subsequent change in the mole fraction of each component in solution. The more volatile components decrease first, and the change is reflected in both the vapor and liquid phase compositions. The composition of the vapor phase thus provides a way to directly monitor the weathering of an ignitable liquid mixture. The composition of the headspace vapors of the containers was monitored by repeated removal of small (c.a. 20 µl) vapor samples from large (c.a. 1 gal.) containers through a septum inserted into the container lid. The dynamic behavior of a mix of volatile hydrocarbons inside a closed container, coupled with vapor adsorption by the portion of the septum exposed to the interior of the container complicated analyte sampling and analysis. The statistical variation in the resulting data did not allow for the determination of slow leaks in the containers. A second method employed in this study was sampling by passive headspace concentration with activated charcoal. This method utilized activated carbon strips to adsorb the hydrocarbon mixture then elute the hydrocarbons from the activated carbon with a solvent. Early experiments by this method were complicated by the preferential adsorption of aromatic and higher molecular weight hydrocarbons by the activated charcoal. Extensive research into the adsorption properties of activated charcoal was conducted1, and the results from the research provided a sampling methodology for monitoring the mole fraction of the C7-C10 hydrocarbon mixture remaining in the container over an extended period of time. A set of four types of quart volume commercial containers held for six different time intervals are undergoing extended time testing. Containers subject to induced leaking are under investigation as positive controls for observable changes in the recovered hydrocarbon mole fraction in the presence of a slow leak. Leak proof systems are under investigation as negative controls against unanticipated changes in the adsorptive function of the activated charcoal over long periods of exposure to hydrocarbon vapors. The results from this long-term study by monitoring hydrocarbon retention in quart volume containers of metal cans, glass jars, and polymer bags will be presented. Data revealing leak rates of the containers and the behavior of the hydrocarbon vapors will be discussed along with the significance to methods in fire debris collection and analysis. References: 1. Mary R. Williams, B.S., Denise Fernandes, B.S., Candice Bridge, B.S., Derek Dorrien, Stefanie Elliott, and Michael Sigman, “Adsorption Saturation and Chromatographic Distortion Effects on Passive Headspace Sampling witth Activated Charcoal in Fire Debris Analysis,” J. Forensic Science, 2004, submitted. Fire Debris, Ignitable Liquids, Trace Evidence 55 B50 Development of a Standardized Field Portable Extraction Gas Chromatography Tandem Mass Spectrometry Method for the Analysis of Ignitable Liquid Residues Marilyn Prieto*, Jeanette M. Perr, BS, Kenneth G. Furton, PhD, and José R. Almirall, PhD, Florida International University, Department of Forensic Chemistry and Biochemistry, 11200 Southwest 8th Street, CP 194, Miami, FL 33199 The goal of this presentation is to discuss the development of a field portable extraction and standardized gas chromatography ion trap tandem mass spectrometry (GC/IT/MS/MS) method for implementation in the detection and identification of ignitable liquid residues (ILR). This presentation will impact the forensic community and/or humanity by increasing the sensitivity and selectivity that is necessary to detect minute amounts of ILR remaining after a fire and to differentiate ILR from IP. It is anticipated that the results using GC/IT/MS/MS will not require sophisticated training, longer analysis times, or data analysis programs. The discovery and recovery of small amounts of ILR in fire debris evidence can be arduous owing to considerable loss of these compounds during the fire, extraction techniques that are not the most sensitive, coextraction of interfering products (IP), and analysis techniques that provide low discrimination. The main purpose of this project was to optimize a general method that can be used by examiners to improve current analysis of fire debris without significantly altering the workload or time required for analysis. In order to accomplish this goal uniform ionization parameters had to be established as well as chromatographic parameters. Uniform ionization regions, called bins (chromatographic references), were created using the standard n-alkanes, C8 through C23, as markers. Alkanes elute from the gas chromatograph in a consequent order depending on boiling point and other factors. The method utilizes an alkane to produce bins that other ignitable liquids (IL) components elute into. The optimized parameters that best fit the IL constituent bin were put forth to evaluate components of interest away from IP thus weakening or even excluding interferences. Distinctive fragments, such as adduct ions, will be utilized to detect the alkane and label the bin. The bin method designed an undiversified and concentrated ionization zone that will result in reproducible mass spectra. Once the optimum MS/MS ionization parameters are selected for each bin, mass spectra will be compiled for IL components. Known IL standards and known weathered IL standards will be tested to generate reference examples using the MS/MS detector through the use of solid phase microextraction (SPME). These acknowledged IL standards will be investigated in the presence of IP, thus exemplifying the advantages, confirm the disadvantages, and establish the utility of this method. This method is expected to increase the sensitivity and selectivity that is necessary to detect minute amounts of ILR remaining after a fire and to differentiate ILR from IP. It is anticipated that the results using GC/IT/MS/MS will not require sophisticated training, longer analysis times, or data analysis programs. Ignitable Liquid Residue, GC/IT/MS/MS, SPME B51 Poking Holes in the Suspect’s Story: An Overview of Pharmaceutical Theft and Tampering Analysis Angela S. Mohrhaus, BS*, Heather A. McCauley, BS, Nicola Ranieri, BS, and John R. Urban, BS, Food and Drug Administration’s Forensic Chemistry Center, 6751 Steger Drive, Cincinnati, OH 45237 The goal of this presentation is to present the forensic community with an overview of the analytical approaches used by the FDA’s Forensic Chemistry Center (FCC) in casework involving the theft of contents from and/or tampering with pharmaceutical vials and syringes. This presentation will impact the forensic community and/or humanity by presenting examples as an analytical roadmap for those in the general forensic chemistry community who may lack experience analyzing similar pharmaceutical fraud/tampering cases. This poster will highlight the approaches used by the FCC in cases involving pharmaceutical theft and/or tampering. The FCC receives suspect narcotic vial and syringe evidence from all over the U.S., and is often asked to answer three basic questions: Has the integrity of the product container been compromised? How much, if any, of the original product remains? What, if anything, was used as a replacement medium (adulterant)? The type of pharmaceutical theft most commonly encountered occurs in hospitals and nursing homes where an individual removes a portion of a narcotic solution from a vial or syringe. In most cases, the individual will “replace” the contents removed with some sort of diluent, such as saline or bacteriostatic water. The FCC has also received cases where the original narcotic product was replaced with another, less potent narcotic. The motive for using a less potent narcotic may be to minimize the chances of the theft being discovered, as a patient receiving the less potent narcotic would be less likely to complain about the pain medication not working. When a suspect syringe or vial arrives at the FCC, its first stop is in the Microscopy Lab. There the vials and syringes are examined for evidence of tampering and determination of what may have been used to access and remove the contents of vials and syringes. Using both Polarized and Stereoscopic Light Microscopy (PLM/SLM), the microscopists are frequently able to determine the number of times, for instance, a vial’s rubber septum has been punctured. Excessive numbers of punctures can be indicative of product theft, adulteration, replacement, or other types of fraud. The shape and characteristics of the puncture hole(s) provides evidence as to what sort of tool was used to puncture the septum. The next step in the process is GC-MS analysis. The contents of the suspect vials and syringes are screened for the presence of the active ingredients, as well as any additional drugs or unusual components. If the contents appear to be diluted, this analysis may indicate what was used as a diluent. For example, if benzyl alcohol (a preservative) is detected in the suspect item using the GC-MS screen, but not the controls, bacteriostatic water or saline may have been used for replacement. After determining the composition of the vial/syringe contents, the compounds present are quantified (if possible) by HPLC with UV detection in order to determine the extent of dilution. The final step, if necessary, in the analysis of these samples is elemental analysis. If during the GC-MS analysis it is determined that the contents were diluted, ICP-AES analysis can be performed to evaluate the contents for the presence of excess sodium relative to untampered control pharmaceutical samples. The amount of excess sodium relative to the dilution of the active pharmaceutical ingredient may be consistent with the replacement of the stolen contents with saline. Once all of the necessary analyses are completed, the pieces of the puzzle start to fit together. Generally, the FCC can report back to the case agent with the following information: how many times the suspect product was accessed, what type of tool was used to access the contents, whether there is active ingredient remaining, whether another drug (or a potentially harmful substance) was used in replacement, how much of each identified component is present in the solution, and whether saline was used as the diluent. Pharmaceuticals, Tampering, Theft 56 * Presenting Author B52 Drug Profile of Urine Specimens From Arrestees in Taiwan Jui Hsu, MSEHS, Wen-Ing Tsay, MS, Chiareiy Liu, M.K. Kuang, MS, Chun-Sheng Chien, and Jih-Heng Li*, National Bureau of Controlled Drugs, Department of Health, #6, Linsen South Road, Taipei, 100, Taiwan Not only in the United State but also in Taiwan, using drugs are criminal behaviors and the drug abusers have the tendency of using polydrug. From this presentation, attendee could learn, retain and even implement into their practice about drug use and profile of urine specimens from arrestees. Drug abuse is a global phenomenon and its extent and characteristics differ from region to region. This presentation describes the most common combinations including MDMA identified were MDMA/ketamine and MDMA/Methamphetamine use in Taiwan, and will impact the forensic community and/or humanity by assisting forensic community to understand the poly-drug combinations. Introduction: Drug abuse is a global phenomenon and its extent and characteristics differ from region to region. Club drugs, MDMA, marijuana and ketamine, have emerged in Taiwan in recent years. MDMA and ketamine positive rates as high as 76% and 47% from rave party participants were shown by Lua et al. in 2001.1 Easy accessibility of precursors and of manufacturing know-how, relatively low prices and the trendy and benign image with popular techno-music and rave culture were all responsible for the spread of MDMA, MDA and MDEA rapidly as reported by UNODCCP in Global Illicit Drug Trends 2000. 2 In Taiwan, using drugs are criminal behaviors and the drug abusers have the tendency of using poly-drug which causes complicated and serious health problems owing to its variety of combination. To understand the drug profile and the combinations of poly-drug use in Taiwan, the authors conducted a study of urine drug testing on drug related arrestees who might commit crime(s) by using drugs. Experiments: 930 urine specimens were collected from police arrestees suspected of drug abuse from twelve of the twenty-three counties in the Taiwan island from Jan 2002 to July 2002. In this study, screened urine specimens were screened with immunoassay reagents adapted to Merck Selectra Vista II and Hitachi 705 automatic biochemistry analyzers according to the manufacturer’s recommendations, then confirmed the presence of morphine, codeine, methamphetamine, amphetamine, ketamine, MDMA and marijuana with GC/MS. At the meantime, REMEDi HS Drug Profiling System was used to screen for the other drugs. Thirtynine drugs were identified positively in 930 specimens. Results and discussion: The REMEDi HS system is widely used in hospital emergency rooms to identify poisons in a patient’s urine or serum. It is highly automated and fast with a potential to screen over 900 drugs and metabolites, including stimulants, local anesthetics, antidepressants, antibiotics and pesticides. In this study, specimens were screened for more than 100 drugs out of the over 200 on Taiwan’s controlled drug list. The drugs include benzodiazepines, barbiturates, synthetic opiates and stimulants. Morphine and methamphetamine were verified as still the major drugs detected in Taiwan, followed by Benzodiazepine (18.3%), MDMA (17.7%) and ketamine (7.2%). Complicated poly-drug use patterns were observed and up to six drugs or metabolites were detected in some specimens which included methamphetamine, benzodiazepine, opiate, precursor, MDMA, MDA and ketamine. The most frequently identified drug combinations were morphine/Methamphetamine (19%), Codeine/Methamphetamine (15%), morphine/amphetamine (15%) and morphine/benzodiazepine (13%). The most common combinations including MDMA identified were MDMA/ketamine and MDMA/Methamphetamine. Conclusion: Morphine and Methamphetamine were verified as still the major drugs of abuse in Taiwan, followed by Benzodiazepines, MDMA and ketamine. Ketamine and benzodiazepines were detected at high rates and became a significant problem. Also for the drug combinations, of * Presenting Author ketamine positive specimens, MDMA was most frequently identified and of benzodiazepines positive specimens, morphine, methamphetamine and opioid were found mostly. The detection rates for THC and barbiturates were low in arrestees. These results revealed important information for further research and policymaking. Reference: 1. Lua, A.C., Lin, H.R., Tseng, Y.T., Hu, A.R. and Yeh, P.C. 2003. Profile of Urine Samples from Participants at Rave Party in Taiwan: Prevalence of Ketamine and MDMA Abuse. Fore. Sci Intl. 136: 47-51. 2. UNODCCP: United Nations Office for Drug Control and Crime Preventions, Studies on Drugs and Crime: Global Illicit Drug Trends, 2000. Arrestee, MDMA, Urine Specimens B53 A Blind Trial Evaluation of a Practical Methodology for Deducing Impact Velocity and Droplet Size From Bloodstains Lee Hulse-Smith, MS*, King’s College London, Strand Campus, London, WC2R 2LS, England; and Mike Illes, Ontario Provincial Police: Forensic Identification, 453 Lansdowne Street, East, Peterborough, Ontario K9J 6X5, Canada After attending this presentation, attendees will learn the blind trial results of a novel technique that utilizes bloodstain diameter and number of spines to derive droplet size and velocity This presentation will impact the forensic community and/or humanity by providing boodstain pattern analysts a methodology that could calculate droplet properties such as size and velocity. Upon the discovery of a bloodstain that meets the criteria (circular with radiating spines), investigators could derive the impact velocity and droplet volume and utilize these variables to: 1. Infer bloodshed forces by correlating impact velocity with an assault instrument. 2. Determine release height of passive droplets impacting on horizontal surfaces. 3. Incorporate projectile motion into trajectory calculations for bloodstains discovered on non-horizontal surfaces. Crime scene reconstruction is dependent on the ability to establish past events from present variables. Investigators now have a method for uncovering droplet properties from bloodstains, leading to a more accurate interpretation of the crime scene. Bloodstain diameter and number of spines are independent variables that can be combined to solve for impact velocity and droplet volume. This was established in a prior experiment that utilized mechanical engineering models to solve droplet properties from pig bloodstains. A blind trial study was subsequently undertaken to test the accuracy of this technique using a redesigned crime scene methodology. Bloodstains found at the scene of a violent crime can often be used to reconstruct the events surrounding bloodshed. However, reconstruction would be enhanced if bloodstain morphology could be used to infer droplet properties. Investigators could then determine the source-of-origin and the forces surrounding bloodshed more accurately. At present, droplet properties are lost upon impact with the surface. As a result, a classification system based on stain diameter has been adopted to dissuade investigators from using bloodstains to estimate droplet properties. Attempts have been made to correlate impact velocity with stain diameter. However, this has proven difficult since the size of a bloodstain is a function of two unknown variables (droplet size and velocity). Alternatively, Balthazard et al. was interested in using bloodstain spines to solve for impact velocity. Spines are commonly found radiating from the periphery of stains, and range in appearance from waves to sharp protrusions. As with stain diameter, Balthazard discovered that spines alone could not predict impact velocity, due to the additional influence of droplet 57 volume. He concluded that a deeper understanding of droplet dynamics was required. Several mechanical engineering models that have since been developed show a predictable relationship between stain morphology (diameter and spines) and droplet properties (size and velocity). In a previous study, Hulse-Smith et al. evaluated two mechanical engineering models for their ability to predict droplet size and velocity from bloodstains. Pig blood droplets were released over a range of diameters (3.0 - 4.3mm) and impact velocities (2.4 – 4.9m/s) onto four surfaces (glass, steel, plastic, paper). The resulting bloodstains were then used to predict droplet properties. A strong correlation was found between observed and expected results. To determine if this technique would translate to the crime scene, a redesigned methodology was tested under blind trial conditions. An investigator was presented with images of human bloodstains and samples of each impact surface. The conditions used to create each bloodstain were not revealed. The investigator subsequently performed a calibration to derive two specific equations for each surface. This aided in eliminating any variation introduced by surface irregularities. Finally, the number of spines and stain diameter were quantified from each unknown bloodstain image and integrated into the respective equations to solve for droplet properties. Experimental results are revealed during this presentation. Bloodstain Pattern Analysis, Blood Droplet, Spines B54 Paradoxical Effects of Surface Structure and Drop Height on Blood Stain Pattern Formation Mark Benecke, MSc, PhD*, and Saskia Reibe, BSc, International Forensic Research & Consulting, Postfach 250411, Cologne, 50520, Germany; Philip von Doetinchem, BSc, Technical University of Aachen, c/o International Forensic Research & Consulting, Postfach 250411, Köln, 50520, Germany; Petra Boehme, BSc, University of Bonn, c/o International Forensic Research & Consulting, Postfach 250411, Köln, 50520, Germany; Andreas Scholten, BSc, and Alexandra Schulz, BSc, University of Cologne, c/o International Forensic Research & Consulting, Postfach 250411, Cologne, 50520, Germany; and Melanie Gericke, KK, Police Academy Hamburg, c/o International Forensic Research & Consulting, Postfach 250411, Cologne, 50520, Germany After attending this presentation, attendees will understand the influence of surface structure on shape of blood drops is very hard to predict without extensive experimentation. Even if surfaces seem to be very similar, paradoxical effects occur. Systematical variation using the actual surface from a crime scene will still allow scientifically sound statements, e.g., relating to height of blood source. Ultra-high-speed photography and physical analysis of the edges of blood stains during their process of formation (Karsten Loehr, Université Pierre et Marie Curie, Paris) also help to understand the dynamics between the surface, the blood drop and its later shape. This presentation will impact the forensic community and/or humanity by demonstrating how extensive experimentation is necessary to understand the influence of the actual environment on the shape of blood drops. Even very reasonable and obvious assumptions about the structure of a surface may easily lead to wrong interpretations, and wrong expert witness statements. During casework, the authors encountered the problem that height of the blood source became relevant. In order to determine the dependence of different types of underground on the shape of the drops, the authors checked the influence of asphalt, paper, linoleum, and plastic against different dropping heights (5, 100, 250, 500, 1000, 2000, and 3000 mm; n (drops) = 520). Then the diameter of the stains was compared, the number of fingers, the maximum distance of satellite drops to the center of a stain, and the total number of satellites. 58 Initially, asphalt and paper produced effects of rough surfaces whereas linoleum and plastic represented rather smooth surfaces. However, the general tendency of the diameter to increase with drop height, and to remain constant beyond a certain height, was not observed on asphalt. Instead, the diameter remained constant irrespective of drop height. Also, the number of fingers (“noses”) of stains produced on asphalt did not increase like on the other surfaces. The largest number of fingers was observed on paper. The maximum distance of satellites to their central stain mostly increased linear with drop height. Asphalt presented the largest distances. Drops on the plastic surface showed constantly small distances of the satellites to the center of the stain. Exact total number of satellites could only be determined on paper and asphalt. Even though both materials showed properties of the rough surface group, they produced very different results (e.g., on asphalt, the number of satellites increases linearly and steeply with height). In contrast, on paper the number of satellites stayed nearly zero. In many cases it also seemed as if the relationship between height and the respective measured parameter was initially not linear but exponential, inverse exponential, or following a saturation curve. These effects will only become visible if small intervals between drop heights are chosen experimentally, especially in the often neglected range between 0 and 100 cm. Conclusion: In casework, extensive experimentation is necessary to understand the influence of the actual environment on the shape of blood drops. Even very reasonable and obvious assumptions about the structure of a surface may easily lead to wrong interpretations, and wrong expert witness statements. Blood Stain Pattern Formation, Crime Scene Reconstruction, Physics of Drop/Surface Interaction B55 ESCRIME: A New Software for Bloodstain Pattern Analysis in 3-Dimensions Philippe Esperanca, MS, and Yves Schuliar, MD*, IRCGN, 1, Boulevard Théophile Sueur, Rosny-sous-Bois, 93110, France; Pascal Chaudeyrac, MSFS, B. Piranda, and Didier Arques, PhD, Université de Marne la Vallée, Section Informatique, cité Descartes Champs-sur-Marne, MarneLa Vallée Cedex 2, 77454, France The goals of this presentation are to explain the possibilities and the use of a software created to reconstruct 3-D trajectories of bloodstains at crime scene. This presentation will impact the forensic community and/or humanity by presenting a new tool to deal with bloodstain pattern analysis at the crime scene. Today, bloodstain pattern analysis is a fully recognized forensic discipline. The shape of stains assists investigators in determining the point of origin when locating the victim and the assailant during the bloodshed. The data obtained are very helpful to confirm witnesses or suspects statements and to support the investigators assumptions. The forensic science laboratory of the French Gendarmerie and Marne La Vallée computer University created a new software called ESCRIME. The function of ESCRIME is to assist investigators with the interpretation of bloodstain patterns by bringing together three functions. The first one is to create a virtual 3-D crime scene according to the real furniture and different textures observed. It is possible to know very precisely (within millimeters) the exact location of each item. The second deals with the exploitation of the pictures of bloodstains. It is important to extract exact information and to measure a stain and locate it at the scene. The third one allows the software to calculate the half plan the stain pass by and to determine point of convergence. The user can easily move inside the crime scene to obtain different views in 3-D. * Presenting Author The authors will present real cases using the software in real time. ESCRIME is not only useful for bloodstain pattern analysts but also for other crime scene specialists, and can help to present cases to the court. Bloodstain Pattern Analysis, Computer Analysis, Crime Scene Reconstruction B56 Identification of Benzimidazolone Organic Pigments in Automotive Coatings Using Pyrolysis Gas ChromatographyMass Spectrometry spectral overlap issues that occur when using FTIR. Py-GCMS also provides information about the polymeric binder or film former. Organic Paint Pigments, Automotive Coatings, Pyrolysis Gas Chromatography-Mass Spectrometry B57 Environmental Effects on Automotive Polyurethane Coatings: Implications for Casework Vincent J. Desiderio, BS*, and David Gardner, BS, New Jersey State Police Office of Forensic Sciences, Central Laboratory, 1200 Negron Road, Hamilton, New Jersey 08691 Janice L. Hellman, MSc, and Gavin R. Edmondstone, BSc*, Centre of Forensic Sciences, 25 Grosvenor Street, Toronto, Ontario M7A 2G8, Canada; and Katrina L. Bernardin, BSc, Forensic Science Unit, University of Strathclyde, 204 George Street, Glasgow, Scotland G1 1XW, United Kingdom This presentation will impact the forensic community and/or humanity by presenting a proposal for an alternate means of identifying benzimidazolone organic pigments in automotive coatings. Automotive coatings frequently play an important role in investigations of vehicular accidents, including “hit and run” incidents. When an automobile hits an individual, another vehicle, or an inanimate object, some portion of the vehicle’s paint is often left behind. This evidence can serve two purposes: 1) If a suspect vehicle is located, a comparison of any questioned paint from the scene or victim to the known paint from the vehicle can be performed; and 2) If a suspect vehicle is not available, any paint left at the scene or on the victim may be useful for developing investigative leads. Of the two possibilities listed above, this paper is primarily concerned with the latter. In order to provide investigative leads, the questioned paint must be examined physically and chemically. Chemical analysis of the paint evidence should be carried out in such a way so that as many of its individual components may be characterized as possible. The results should then be compared to a comprehensive database to determine whether a possible make, model, and year of suspect vehicle can be ascertained. The collaborative efforts of the Royal Canadian Mounted Police (RCMP) and FBI (FBI) have provided the forensic science community with such a database in the form of the Paint Data Query (PDQ). The PDQ is based on the input of data obtained from visual, elemental and spectroscopic analysis of questioned samples. Generally, as more components of questioned paints are identified, the discrimination potential of such evidence increases. With recent shifts away from inorganic pigments, which often contain heavy metals such as Pb and Cd, organic pigments have become more prevalent in automotive coatings. Therefore the identification of organic pigments would be advantageous in generating a shorter list of possible suspect vehicles. In its current state, the PDQ does not include organic pigments in its identification scheme. This absence may be due in part to a lack of research in this area. It has recently been shown that organic pigments can be identified in automotive paints using Fourier Transform Infrared Spectroscopy (FTIR). However, in certain instances the relatively low concentrations of organic pigments typically found in automotive coatings may make it difficult to make an identification using FTIR alone. In addition to the problem of low concentrations of organic pigments, automotive coatings frequently contain inorganic pigments, flakes and fillers that tend to obscure the presence of the lower levels of organic pigments when using FTIR. To address this problem, this research was undertaken to evaluate the use of Pyrolysis-Gas Chromatography-Mass Spectrometry (Py-GCMS) as a compliment to FTIR for the identification of organic pigments. PyGCMS can provide lower detection limits so that certain problematic organic pigments may be conclusively identified. This technique also provides for a cleaner separation of the contributions of the organic components from those of the inorganic components, thus offsetting the The goal of this presentation is to present the infrared spectral variability that can occur within the same automotive polyurethane clearcoat to promote a better understanding of how this can influence casework for paint examiners. This presentation will impact the forensic community and/or humanity by showing paint examiners why it is important to fully characterize polyurethane clearcoats when conducting a comparison analysis of this material. In particular it is important to sample at various depths from the surface or at least to sample at a consistent depth from the surface. Without proper consideration of the internal variability of these coatings incorrect exclusions could be made. Chemical variability within automotive polyurethane clear coats has been observed in casework at the Centre of Forensic Sciences. These chemical variations were observed in the infrared spectra as differences in the relative intensity of the prominent band at 1690 cm-1 with respect to the 1730 cm-1 carbonyl band. The differences in intensity appeared to be dependent on the depth from the surface in automotive paints. A common technique of sampling clear coats for IR analysis is to manually slice thin peels, a method that does not permit the depth of sampling to be carefully controlled. This could contribute to the variation, and cause difficulties in interpreting a paint comparison. It has been documented in the literature that polyurethane paints degrade upon prolonged exposure to UV radiation and water. The chemical changes due to the photo oxidation of these coatings have been studied using infrared and UV spectroscopy. However, in these instances the degradation process was initiated using weathering simulations. It is not known to what extent this degradation occurs under normal exposure to the environment and whether it would vary depending on the location on the vehicle. It has also been reported that the presence of water vapor during the curing process can create depth dependant variability in the polyurethane film. This may also account for the spectral variations observed. Determining the extent of chemical variation that can occur within polyurethane automotive clear coats of real automobiles (not test panels) would be useful for interpreting these variations when they occur in casework and for developing sampling strategies. Polyurethane clear coats are used extensively as automotive repaints and to a lesser extent as OEM (Original Equipment Manufacture) clear coats, thus they are frequently encountered in cases that require analysis of automotive paint. Differences of the relative intensities of infrared bands that appear to be reproducible could be interpreted as a differentiating feature between two paints. However, if these differences are due to internal variability of the coating that is not adequately represented then it is not a true difference. Thus, variation within the polyurethane clear coats could cause problems when interpreting infrared data especially when working with very small samples. Polyurethane clear coats were obtained from a variety of automobiles and were analyzed using Fourier transform infrared spectroscopy to * Presenting Author 59 determine the chemical variations within these paints. Chemical changes in the polyurethane clear coats were assessed as a function of depth (distance from the surface) by the examination of cross-sectioned samples by infrared micospectroscopy. Differences over different locations on the automobile were examined using attenuated total reflectance spectroscopy, which is a surface sensitive technique. Distinct chemical variations were observed both as a function of depth, and of location on the vehicle in a significant minority of the samples investigated. These differences could lead to an erroneous interpretation. While demonstrating the spectral variability within a clear coat is time consuming it can be essential to achieving a correct interpretation. The data from this analysis will be presented and the implications of these findings on data interpretation and sample preparation will be discussed. Automotive Paint, Polyurethane, FT-IR B58 Characterization of Automobile Body Fillers Jay A. Siegel, PhD, Indiana University, Purdue University, School of Science, 402 North Blackford Street, Indianapolis, IN 46278; Sara McNorton, MS*, 2544 Chilver Road, Windsor, Ontario N8W 2V9, Canada; and Guy Nutter, BS, Michigan State Police, 42145 West Seven Mile Road, Northville, MI 48167 After attending this presentation, attendees will understand the characterization of auto body fillers by instrumental means. This presentation will impact the forensic community and/or humanity by providing forensic scientists a mean to compare and characterize body fillers will be greatly increased. The objective of this study was to determine if chemical and physical differences in body fillers from various manufacturers exist and could be identified. Extensive research was done to obtain a representative sample of automobile body fillers and spot putties used in the U.S. After all of the samples were identified, 33 samples were obtained. Twenty-four of the samples were body fillers and 9 were spot putties. Nine different companies manufactured the 33 samples. All of the samples were analyzed by FTIR microscopy, stereo microscopy and pyrolysis GC-MS. The results of the analysis showed that 13 of the 33 samples could be correctly identified as to source, while the rest of the samples could be put into groups containing 2-4 types. Automobile, Body Fillers, Spot Putties B59 Beyond the Barcode: The Electronic Product Code and the Future of Product Identification Max M. Houck, MA*, 886 Chestnut Ridge Road, PO Box 6216, Morgantown, WV 26506-6216 Attendees will learn about the electronic product code (EPC), the object naming system (ONS), and how the coming tide of product identification technology, such as radio frequency identification (RFID), will affect their ability to identify products and track manufacturers. This presentation will impact the forensic community and/or humanity by providing forensic scientsts with an understanding of the coming systems of product identification, such as the EPC and the ONS, to better identify products that are submitted as evidence and track the product to its manufacturer or even its point of sale. Without an awareness of this information, crucial evidence to a case may be neglected or lost. Just as the UPC code has transformed retail operations around the globe over the past twenty years by increasing productivity and efficiency within the supply chain, the EPC (electronic product code) could take 60 supply chain dynamics to the next level. The EPC is a new product numbering standard under development by the Uniform Code Council (UCC) that can be used to identify a variety of items using radio frequency identification (RFID) technology. The 96-bit EPC code links to an online database, providing a secure way of sharing product-specific information along the supply chain. Like other RFID solutions, the EPC’s ability to be read without a line-of-sight offers users significant savings to manufacturers and retailers. The existing UPC code allows retailers to track products at the SKU (stock keeping unit) level by providing every product with a unique identifier. The UPC (and its International cousins, the EAN and JAN) is made up of two key components: a number which identifies the manufacturer of the product, and a number which identifies the product belonging to that manufacturer. Each time a new product is created, or an existing product is modified in any way (including changes in packaging), a new UPC code is assigned to the product. Since each product may go though several minor design/packaging changes over its life, a single product may end-up with several UPC codes that identify it to a retailer, even though the retailer may consider the product as a single SKU. The EPC technology, in conjunction with the expanding production of RFID capable printers/encoders, has the potential to revolutionize the supply chain by providing more accurate information about product movement, stock rotation, inventory levels and other management information. It also would be a significant tool for product recalls and theft prevention. A consortium of companies (including WalMart, Proctor and Gamble, and Sun) is currently supporting research into this new technology. The EPC code is a new product numbering standard that goes way beyond identifying products. The EPC assigns a unique number to every single item that rolls off a manufacturing line—that is, every single bottle of soda would have its own unique EPC number. The EPC will allow every company in the supply chain, including retailers, to track products at the individual item level. This means every single item on a shelf could be traced back to when it was made and when it is sold. However, the structure of the EPC does not necessitate every retailer track items at this level. The EPC has been designed to allow it to replace the UPC and allow tracking at the SKU level if desired. Because of the enormous quantity of unique numbers required to track at the item level, the EPC utilizes a “96-bit numbering scheme.” The implications to forensic science are enormous—the ability to track products that appear as evidence back to their point of sale! It is critically important that forensic scientists become aware of this new technology, how it works, and what resources are available to keep tabs on this groundbreaking technology. Barcodes, Product Identification, Electronic Product Codes B60 Forensic Science and Homeland Security: A Critical Piece of the Puzzle Barry A.J. Fisher, MS, MBA*, Scientific Services Bureau, Los Angeles County Sheriff’s Department, 2020 West Beverly Boulevard, Los Angeles, CA 90057 State and local crime labs, for the most part, are unprepared to provide assistance in terrorism acts or mass casualty incidents. This presentation will impact the forensic community and/or humanity by providing attendees with a clearer understanding that State and local crime labs are not part of a nation preparedness strategy. Attendees will also receive suggestions on how to remedy this situation in their home regions. Suppose terrorists mount coordinated attacks across the United States and abroad striking many targets simultaneously. Historically, the FBI, FEMA, D-MORT teams, and other organizations provide assistance to local agencies during disasters with or without mass casualties. But what * Presenting Author would happen if the federal response were stretched so thin that local jurisdictions were on their own for a few days or longer? Naturally, the “locals” would fend for themselves and manage as best as they could. A serious problem exists today. There is no defined role for State and local crime labs who employ thousands of trained scientists throughout the United States in the event of a terrorist incident. Despite their wide use in other areas, local crime labs have largely been ignored as an added resource for homeland security related incidents. The result is that few state or local crime labs are prepared to help out should an incident occur. The idea that forensic science personnel might be among the first responders or support first responders has not been fully considered. Developing a local forensic science response to a terrorist incident while it is unfolding is not the appropriate time to start planning. While crime labs do not have the capacity to specifically handle certain types terrorist incidents as certain Federal agencies they, none-theless, in a WMD scenario, they will play a role, whether they are prepared or not. The events in New York City following September 11, 2001, demonstrate this when the NYC Office of the Chief Medical Examiner conducted thousands of DNA tests for victim identification. State and local public safety personnel will be involved with pre and post terrorist events. It is no leap to expect local forensic science personnel to also be involved as either first responders or immediately following the first responders. They could be involved at the scene and in the laboratory as examiners of physical evidence in traditional forensic science efforts such as, DNA testing, latent fingerprint examination, handwriting examination, and so on. Before forensic scientist show up at a scene or become involved, careful consideration will be given concerning chemical, biological, radiological and nuclear contamination of evidence, analysis of large volumes of evidence, tracking evidence, coordinating information, training and preparedness. To date, federal planners that would ultimately manage terrorist incidents have not considered what might be the appropriate role for State and local forensic science laboratories in terrorist incidents. Other than medical examiners and coroners, there have there been few attempts to prepare local forensic practitioners for this assignment. What are appropriate steps that might be taken to remedy the present situation? Just as the Department of Health and Human Services has brought state and local public health labs together under a “laboratory response network” an appropriate federal agency should bring together state and local crime labs along with homeland security stakeholders to develop a role for state and local forensic science laboratories and begin planning how to use these resources. A national forum is one way to define the relationship of forensic science to homeland security. Some of the issues that might to be discussed include: the identification of mass casualties by DNA; the examining WMD contaminated evidence at the crime lab; the coordination of local crime labs with state and local public health labs; training and equipping forensic science personnel; and integrating forensic science into WMD preparations. State and local crime labs can also begin preparations now by contacting local stakeholders concerned with terrorism and mass casualty planning. Sometimes it takes the prodding of the “locals” to get the “fed” moving. And now would certainly be a good time to begin, before the next incident occurs. Terrorism, Homeland Security, Mass Casualties * Presenting Author B61 Bioterrorism in Europe After September 11th: Our Experience in BIOTOX Procedure Thierry W. Faict*, and Stéphanie W. Neel, MD, Faculté de Médecine, CHU de Clermont-Ferrand, 28, Place Henri DUNANT, ClermontFerrand Cedex 01, 63001, France This presentation will impact the forensic community and/or humanity by showing the forensic community that forensic scientists should be involved in bioterrorism for many reasons among which are the protection of the population of penal evidences At the end of 2001, the United States of America was hit by incredible acts of terrorism. In the months and years following, small groups have tried to increase the stressful climate by using militarized “harmless” bacterial strain. The effects of globalization allowed a wild distribution, not of the bacterium, but of fear, especially in Europe and analysis on different supports suspect to be infected are being perfomed today. The authors are showing here the impact and the experience of a forensic ward in the fight against bio terrorism. Systematic bacteriological and judiciary analysis of suspect postal dispatches in France from October 23, 2001 to July 2004 are exposed. Bacteriological and toxicological results, as well as penal consequences for the authors have been studied. The administration sought to implement this procedure for sanitary and economical reasons and for penalty proof protection as well. More than 250 analyses were performed and no anthrax was found. The organization of this procedure, its history and difficulties are presented. The techniques and bacteriological methods used will be analyzed. Methods implemented in order to preserve judicial proof are also presented along with the difficulties in relation to their qualification for efficient penal procedures. Bioterrorism, Anthrax, Europe B62 Census of Publicly Funded Crime Laboratories - 2002 Joseph L. Peterson, DCrim*, Department of Criminal Justice, University of Illinois at Chicago, 1007 West Harrison Street, Chicago, IL 60607; and Matthew J. Hickman, MA, Bureau of Justice Statistics, 810 Seventh Street, NW, Washington, DC 20531 By attending this presentation, attendees will receive an overview of a recently completed census of the nation’s publicly funded crime laboratories. This presentation will impact the forensic community and/or humanity by providing forensic laboratories and policy makers with crime laboratorie’s staffing patterns and caseload characteristics, as well as their resource needs. This presentation will provide laboratories and policy makers with crime laboratories’ staffing patterns and caseload characteristics, as well as their resources needs. The Department of Criminal Justice at the University of Illinois at Chicago (UIC), in collaboration with the American Society of Crime Laboratory Directors (ASCLD), was funded by the Bureau of Justice Statistics (BJS) to undertake a census of all publicly funded crime laboratories throughout the United States. Because this was to be a census, responses were sought from every, individual crime laboratory in the nation. UIC was asked to gather baseline statistical information for 2002 on the operations and workload of crime laboratories, and to assess where added resources were needed. The objective was to improve public understanding of the work performed and resources committed to crime laboratories. 61 Staff worked extensively with ASCLD and the crime lab community to develop a comprehensive listing of laboratories. The survey itself had six primary areas: Organization, Budget, Staff, Workload, Outsourcing, and Quality, Training, & Research. First the governmental entity and population of jurisdictions served was determined, and types of forensic functions performed in each laboratory. Next, the budget breakdown of each laboratory was examined, and source of funding. We then determined laboratories’ workloads and queried labs about the number of cases received and backlogged, and the volume of requests received, worked and backlogged in 2002. Also determined was the performance expectations that laboratories had of various examiner specialists, and what added resources would be needed to reduce backlogs. Questions were asked about the number of requests outsourced, as well as the cost and funding sources for such testing. Lastly, laboratory accreditation was determined, the types of proficiency tests in which they were engaged, and the dedication of resources to training and research. Facilities had the option to respond either via paper or electronic form. The process began with a mailing list of 469 facilities that were queried to ascertain if they met the definition of a publicly funded crime laboratory. Ultimately, 126 entities were excluded that were either duplicates, bad addresses, or did not meet the definition of a laboratory; e.g., many crime scene and police ID units were excluded. Three hundred forty three (343) laboratories remained on the list as bona fide crime laboratories. Once it was determined that all survey responses from all laboratories willing to complete the primary survey were received, an abbreviated (short) survey was developed abd directed to the remaining, nonresponding laboratories. Two hundred eighty-six 286 full and 25 short (partial) surveys were received from participants – a total of 311 individual laboratory responses, representing a response rate of about 90%. Also received were 15 aggregate responses representing multiple laboratories in state or federal systems. Data were initially compiled and published via a BJS Fact Sheet for the fifty largest state and local laboratories in the United States. These facilities employed more than 4,300 full-time equivalent (FTE) personnel in 2002 and had budgets exceeding $266 million. Laboratories received almost one million cases in that year, including over 1.2 million requests for forensic services. The requests represented about half of all requests for forensic services handled by publicly funded laboratories nationally. These labs ended the year with over 93,000 backlogged cases including about 270,000 requests for forensic services – more than twice the backlog that existed at the beginning of the year. It was DNA analysis were backlog was increasing the fastest; i.e., for every DNA analysis request completed in these laboratories in 2002, an estimated two requests were added or remained outstanding at yearend. A summary of all data collected will be presented at the meeting. Crime Laboratories, Survey, Backlog B63 Biotracks – Leveraging DNA Technology to Solve Lesser Offense Cases and Reduce Recidivism in Queens County, New York Denis McCarthy, MA, MBA, W. Mark Dale, MBA, and Lisa Faber, MFS*, New York Police Department Laboratory, 150-14 Jamaica Avenue, Jamaica, NY 11432; Charles S. Hirsch, MD, Office of the Associate Chief Medical Examiner, 520 First Avenue, New York, NY 10016; Mitchell M. Holland, PhD, and Michael Cariola, MFS, The Bode Technology Group, 7364 Steel Mill Drive, Springfield, VA 22150; and Charlotte J. Word, PhD, and Mark D. Stolorow, MS, MBA, Orchid Cellmark, 20271 Goldenrod Lane, Germantown, MD 20876 The objective of this presentation is to demonstrate the value of recognizing and collecting various types of biological evidence left at burglary scenes. By generating STR profiles from evidence such as perspiration, hair, and blood, and uploading them into local, state, and federal 62 DNA databases, the perpetrator(s) of a no-suspect case may be identified and links between otherwise unrelated burglaries may be established. By maximizing the potential and effectiveness of the DNA databases, this study demonstrates a means of identifying the recidivism rates of convicted offenders within a specific geographical region, as well as identifying any correlation between burglaries and more violent crimes such as sexual assault and homicide. Research has shown that virtually all offenders begin their criminal careers early in life with lesser offenses. A 2002 study by the Bureau of Justice Statistics shows that burglars had one of the highest re-arrest rates at 70.4% as well as one of the highest recidivism rates for specialists i.e., prisoners who, after being charged with one type of crime, are likely to commit the same type of crime again. Considering that there are over 110,000 convicted offender DNA samples in the New York State database to date, the authors felt it was time to leverage this technology and focus on lesser offenses such as burglary in order to reduce their frequency and ultimately reduce the frequency of more violent crimes. In order to ensure that the most probative items of biological evidence are collected and analyzed, it is essential to have a well-trained and informed evidence collection team. In addition to targeting conventional items such as cigarette butts, blood, soda cans, and hair, the burglary scene processors in Queens County, New York were trained to recognize and swab contact surfaces such as windows, knife handles, and door knobs which may contain trace amounts of biological material left behind by intruders. Instruction in the protection of these biological samples from contamination, loss, or deleterious change was a key aspect in the training. This presentation will conclude by discussing which types of biological evidence and contact surfaces yielded the most informative STR profiles and how many profiles were developed from specific types of items. With this type of data, law enforcement will gain a better understanding of which items from a burglary scene are likely to yield probative profiles and how many samples need to be collected and analyzed. By obtaining STR profiles from biological evidence left at burglary scenes and utilizing the expanding DNA databases of convicted offenders, the rate of burglaries and more violent crimes can ultimately be reduced. Burglary, DNA, Database B64 Development of Improved DNA Extraction Method for Detection of Biological Threat Agents Collected on Environmental Swabs Richard Winegar, PhD*, Coridon Quinn, BS, Oscar Ruiz, PhD, and Jennifer Wolack, BS, Midwest Research Institute, 1470 Treeland Boulevard, Palm Bay, FL 32909; Douglas Anders, PhD, FBI Hazardous Materials Response Unit, 2501 Investigation Parkway, Quantico, VA 22135; and Kevin W.P. Miller, PhD, FBI Laboratory, Counterterrorism Forensic Science Research Unit, Building 12, Quantico, VA 22135 Attendees will learn an improved method for recovering and detecting DNA from biological threat agents collected on environmental swabs. This presentation will impact the forensic community and/or humanity by demonstrating that sensitive detection can be achieved by using an integrated approach for sample processing that considers recovery of agent from the sampling matrix, release of DNA, counteracting environmental inhibitors, concentration of extract, and robust PCR conditions. Investigations of crimes involving dangerous biological agents often require the use of swabs and wipes to recover the biological evidence. Current laboratory protocols for rapid extraction and detection of Bacillus anthracis (BA) from environmental swabs often rely on the recovery of DNA present on the external surface of spores. Furthermore, these methods generally do not include ways to counteract environmental PCR inhibitors except for dilution of the original DNA sample. As a result, these methods can suffer from poor sensitivity of detection. Additionally, such methods for extraction of spore DNA may not be appropriate for extraction * Presenting Author of nucleic acids from other biological threat agents such as viruses. This study was conducted to determine whether the sensitivity of detection can be increased by incorporating steps for lysis, DNA purification, and optimized PCR conditions. An extraction method was developed which consists of placing a swab head inoculated with microorganisms into a 2 ml O-ring screw-cap tube containing lysis buffer, glass beads and polyvinylpolypyrrolidone. The microorganisms were lysed by bead-beating and the DNA extracted and purified using the Qiagen DNeasy kit. To counteract the effect of environmental PCR inhibitors, molecular biology-grade bovine serum albumin (BSA; 200 ng/mL) was included in the PCR reaction. In order to evaluate this method for extraction of nucleic acids from various types of microorganisms, real-time PCR assays were developed for the detection of DNA from BA (a Gram positive bacterium), Yersinia pestis (YP; a Gram negative bacterium), Vaccinia virus (VAC; an enveloped virus) and Adenovirus (ADV; an un-enveloped virus). Using this extraction method, the reliable lower limit (10/10 extraction replicates showing positive results in duplicate PCR reactions) of detection by realtime PCR is 100 colony forming units (cfu) of BA spores and 100 plaque forming units (pfu) of Vaccinia virus inoculated directly onto clean, sterile swabs. In order to mimic dirty or dusty environmental samples, swabs were coated with 10 mg of urban particulate matter (NIST SRM 1648) prior to inoculation with microorganisms. Real-time PCR detection following DNA extraction from these dirty samples resulted in reliable lower limits of detection of 500 cfu BA and 100 pfu VAC. Despite the apparent poorer detection limit of BA in dust-coated swabs, 100 cfu could be detected in a majority (95% of extraction replicates and 90% of PCR replicates) of the samples tested. Similar studies are underway to determine the limits of detection for YP and ADV. In conclusion, the authors have developed a method for the efficient extraction and sensitive detection of BA and VAC DNA from environmental swabs. Future studies will focus on the recovery and detection of other organisms and development of extraction methods for other sampling matrices. consists of a sample digestion step followed by purification and concentration with a microcon 100 (Millipore). Moreover, this procedure can be automated through the robotic removal of the digested DNA from the swab and transfer to the microcons, assembled into a 96 well plate, the Microcon 96 Retentate® Assembly Plate (Millipore). The Dacron® and cotton swabs tended to clog the pores of microcons hindering sample concentration. Moreover, the microcon elution volumes were inconsistent, often unnecessarily diluting the samples. Since the microfiber also had some problems with liquid retention, additional candidate fibers were selected based on their high absorption potential coupled with the likelihood that their structure promoted fluid release. A series of five analogous, natural fabrics (A-E) were evaluated for potential use for the LCN swab. Fabrics A and B have similar shapes, but had few variations in their weaves, whereas fabrics C, D and E had different shapes, but had few variations in their weaves. To test absorbance, the fabrics were immersed in 10 uL of control DNA in a microfuge tube. When the tube appeared dry, the fabrics were discarded and water was added to the tubes to resuspend any DNA left behind. In order to test the release of DNA, control DNA was deposited directly onto the fabrics, and allowed to be absorbed into the fabric for ten minutes. The fabric was then placed in sterile water and shaken at room temperature for 20 minutes. Following sample concentration, DNA was measured and the percent recovery was calculated. Based on the results, Fabric B and Fabric C were superior. To confirm this finding, fingerprints were collected from volunteers, and were swabbed with each of the two fabrics. Comparable yields resulted. Subsequently, the fabrics were compared with respect to their performance with the robotic system. Prior to this testing, the optimal length of the fabrics for DNA recovery was determined, although, regarding robotics, the smallest length possible is best. Lengths of 1 cm, 2 cm and 3 cm were used to swab dried cells from a surface and extracted. Fortunately, the shorter lengths of fabrics yielded the most DNA. Therefore, the sample digests were removed from Fabrics B and C on the Biomek 2000. However, Fabric B caused a malfunction whereas Fabric C was compatible with the process, and was selected as the candidate swab fabric. Microbial Forensics, DNA Extraction, Biological Threat Agents LCN samples, DNA, Automation B65 B66 Evaluation of Fabrics for Development of a Low Copy Number Sampling Swab James A. Sebestyen, BS*, Mechthild Prinz, PhD, Robert Shaler, PhD, and Theresa A. Caragine, PhD, Office of Chief Medical Examiner, 520 First Avenue, New York, NY 10016 The goal of this presentation is to compare different fabrics that were tested in the development of a swab for the High Sensitivity Laboratory. This presentation will impact the forensic community and/or humanity by developing a swab to improve recovery of DNA from Low Copy Number DNA samples such as fingerprints. The shape of this swab also accommodates robotics, and thus promotes the processing of large numbers of evidentiary samples. In order to maximize the recovery of DNA from surfaces potentially containing Low Copy Number (LCN) DNA samples, such as fingerprints, several fabrics were evaluated based on their ability to absorb and subsequently release cells. These fabrics were also compared regarding their compatibility with the laboratory’s high throughput system. Studies conducted previously tested a wide variety of commonly available fabrics including but not limited to cotton, polyester, and microfiber, and Dacron® and cotton swabs, which are currently utilized for sample collection. Initially, microfibers recovered the most DNA from fingerprints. Cotton and Dacron® absorb liquid very well, and their poor DNA yield from LCN samples suggests that they trap DNA and liquid within their fibers. The Dacron® and the cotton swabs were also not compatible with the optimized extraction protocol, unlike the microfiber swab. This procedure * Presenting Author Effect of Lubricants and Nonoxynol-9 Exposure on Biological Evidence From Condom Ginger Luccero, MFS, and Ismail M. Sebetan, MD, PhD*, Forensic Science Program, National University, 11255 North Torrey Pines Road, La Jolla, CA 92037 Attendees will further their knowledge when dealing with biological evidence exposed to lubricants and spermicides due to use of condom in sexual assault cases. The presented information will provide a starting point for further research on utilization of the rate at which nonoxynol-9 degrades the sperm cell membrane to help in determination of time of incident. This presentation will impact the forensic community and/or humanity by demonstrating research to those working in the field that will further their knowledge when dealing with biological evidence exposed to lubricants and spermicides due to use of condom in sexual assault cases. The presented information will provide a starting point for further research on utilization of the rate at which nonoxynol-9 degrades the sperm cell membrane to help in determination of time of incident. Condom use during sexual assaults has increased and, as a result, so has the forensic significance of condom-associated biological samples. Preservation of this valuable evidence has prompted the investigation of condom lubricants and spermicides for potential degradation of biological samples. Integrity of samples can be inferred by the ability to amplify short tandem repeats (STRs) on the Y chromosome. Y chromosome STRs are male-specific and are polymorphic in the number of times a sequence motif is repeated. As a result, Y chromosome STRs provide a great power of 63 discrimination among individuals. It is important to prove that lubricants and spermicides from condoms do not decrease the accuracy of DNA profiles, and therefore, do not undermine the validity of such evidence in court. Semen samples were incubated at room temperature and at 37° C for up to three days in the presence of a water-based lubricant, an oil- based lubricant, and the spermicide nonoxynol-9. Untreated samples were also investigated for comparison. Samples were harvested for DNA, which served as a template for PCR and quantitative PCR (qPCR). PCR targets included DYS 385, DYS 389I, and DYS 393 loci. For studying rate of degradation of the sperm cell membrane, Semen samples were incubated in condoms in the presence and absence of nonoxynol-9. Nonoxynol-9 (N-9) is classified as a nonionic surfactant and interacts with lipoproteins of cell membranes. Through a time course, samples were fixed with 4% paraformaldehyde and stained with propidium iodine (PI). Cells were sorted on the Guava PCA cell sorter and the percentage of PI negative cells (viable cells) and the percentage of PI positive cells (non-viable cells) were calculated. Successful amplification of all DNA samples was demonstrated through qPCR analysis as well as gel electrophoresis of PCR products. Results also indicated that, over time up to three days, the percentage of viable cells decreased in N-9 treated samples but stayed constant in untreated samples. In conclusion, it has been proven that exposure of semen samples to oil-based lubricants, water-based lubricants, and nonoxynol-9 does not prevent successful Y-STR. In addition, by determining the percentage of viable cells in a sperm sample, the length of exposure to N-9 can be estimated DNA, Y-STR loci, Lubricants & Spermicides B67 Optimization of DNA Extraction Procedures for Low Copy Number Degraded Samples Taylor M. Dickerson, MS*, Mechthild Prinz, PhD, Robert C. Shaler, PhD, and Theresa A. Caragine, PhD, Office of Chief Medical Examiner, 520 First Avenue, New York, NY 10016 Attendees will learn modified versions of common DNA extraction procedures that can be used for low copy number degraded samples This presentation will impact the forensic community and/or humanity by providing optimized procedures to ensure that a high yield of DNA is obtained when a LCN degraded sample is encountered. Ultimately, a higher yield of DNA from the extraction will provide more DNA for amplification. As LCN degraded samples are encountered more frequently in forensic casework, it would be useful to optimize current DNA extraction procedures in order to obtain a higher yield of DNA. The following DNA extraction procedures were investigated: DNA IQ (Promega), QIAamp (Qiagen), 5% Chelex® (BioRad), and a LCN extraction procedure developed at the Office of Chief Medical Examiner with the use of 0.01% SDS. Initial experiments were performed with purified DNA in amounts of 100 pg or less. The DNA was degraded with DNase I and subjected to each of the four extraction procedures, before and after optimization. Human embryonic kidney cells were also degraded with DNase I; and known amounts (~20 cells) were used in each of the extraction procedures. Bloodstains that were degraded with irradiation were also used in this study. As a control, LCN undegraded samples of the same nature were subjected to each extraction procedure, before and after optimization. All of the samples were quantitated with real-time PCR, amplified with the Power Plex 16® (Promega) multiplex system, and separated with the 3100 Genetic Prism® Analyzer (ABI). For the DNA IQ (Promega) extraction procedure, the following parameters were changed to optimize the extraction of the LCN degraded samples: smaller volumes of resin and lysis buffer, shorter incubation time, and the addition of Poly A RNA. For the QIAamp (Qiagen) extraction 64 procedure, the addition of Poly A RNA and an increased temperature during the elution step optimized the extraction. The extraction procedures with 5% Chelex® and 0.01% SDS both gave optimal results with shorter incubation times. DNA yield, or percentage recovery, was calculated for each extraction. The results from the optimized procedures were compared to those before optimization. An increase in DNA yield was seen with all of the optimized procedures. Usable DNA profiles were obtained for all of the samples subjected to the optimized procedures. DNA Extraction, Low Copy Number, Degraded DNA B68 DNA Extraction of Archived Palm Prints: Implications for Cold Case Evidence Jason Berger, MS*, Mechthild Prinz, PhD, Robert C. Shaler, PhD, and Theresa A. Caragine, PhD, Office of Chief Medical Examiner, 520 First Ave, New York, NY 10016 Attendees will learn how DNA can be extracted from archived palm prints, allowing for the genereation of CODIS compatible PCR DNA profiles. DNA considerations for the collection of palm print evidence. This presentation will impact the forensic community and/or humanity by showing how archived palm prints, which may be too smudged or incomplete for traditional fingerprint identification, can be used for PCR DNA typing. This method could allow for the examination of cold cases in which this type of evidence is present. Obtaining a CODIS profile could allow for another way to identify a suspect. The goalof this presentation is to improve the recovery of DNA and the production of PCR DNA profiles from palm prints that have been dusted with soot based fingerprint powder, and archived by tape lifting. Archived latent prints are often the only probative evidence in many unsolved cases. Employing protocols previously developed for Low Copy Number (LCN) DNA samples by the High Sensitivity Team in the laboratory of the OCME, the DNA recovered from tape lifts of fingerprints was often insufficient to produce DNA profiles that could be compared to the national database, CODIS. Therefore the extraction method was optimized and applied to archived palm print evidence, in order to maximize DNA recovery. Samples were collected from ten different subjects on two types of surface (Linoleum tiles and Plastic). Subjects, at least one hour after washing their hands, were asked to press both of their palms onto each surface for five seconds. The prints were then dusted using both black and dual-use fingerprint powders. Following enhancement, the prints were lifted from the surface using a clear adhesive tape and stored on a standard fingerprint lift card. In order to diminish the adhesive properties of the tape, the cards were stored in the freezer at -20° C for at least four hours prior to extraction. The tape was then removed from the card to allow the digestion buffer, 5 mls of 0.01% SDS and Proteinase K, access to the epithelial cells attached to the tape. Thirty minutes was sufficient to digest the cells, yet still preserve the DNA. Samples were purified and concentrated with a centriplus 100 (Millipore) followed by a microcon 100 (Millipore) with the addition of Poly A RNA to prevent loss of DNA. DNA recovery was measured with SYBR Green I and real time amplification of an ALU sequence. All samples containing at least 6 pg of DNA were amplified for 35 cycles with PowerPlex® 16 (Promega) PCR reagents with a halved reaction volume and a doubled extension time. 4 µL of each PCR product was injected at 3 kV for 20 seconds on the ABI 3100 Genetic Prism® Analyzer. Data is expressed as the percentage of correctly determined alleles out of the possible thirty-two alleles amplified. Twenty eight palm prints were collected and archived from six different subjects. At least 6 pg of DNA was recovered from 64.3% of these samples. This DNA was amplified to produce allelic determinations that were 57.4% correct. However, many of these samples were multi-component mixtures. Since extraction of the tape alone produced DNA alleles, * Presenting Author the source of this contamination could be due to prior handling of the adhesive edges of the rolls of tape. Therefore, the middle of the tape containing the print was excised and extracted. Control pieces from the center of the tape did not contain extraneous sources of DNA. Consequently, fifty more palm prints were processed with two methodologies to accommodate contamination from the tape. Half of the prints were collected as described above, but only the middle portions of the tape lifts were digested. Alternatively, the remaining prints were collected on tape that was UV treated prior to sample collection. Similar to the previous study, 65% of these samples contained at least 6 pg of DNA. However, these experiments demonstrated a significant decrease in the number of drop-ins. The non UV-treated tape averaged 46.34% correct alleles with 5.3 spurious alleles called, and the UV-treated tape produced on average 52.2% correct alleles with 15.2 spurious alleles. Furthermore, in order to generate reliable profiles, samples were amplified twice, and alleles were assigned only if they were occurred in both amplifications. Employing only concurrent correct allelic determinations, usable database eligible profiles were apparent, on average, in thirty percent of the samples examined. These studies suggest that archived palm and likely fingerprints may provide DNA evidence. In order to avoid sources of contamination, collection tape could be treated with UV and the DNA analyst should only process the center of the tape. Regarding archived prints from cold cases, the latter strategy is feasible, but depending on collection precautions, mixtures should be anticipated. Low Copy Number DNA, Tape Lifts, Finger/Palm Prints B69 Select Agent Microbial Forensic DNA Identification Techniques Susan W. Jones, PhD*, Richard P. Schoske, PhD, Stephen Francesconi, PhD, Lynn Cooper, PhD, Michael Dobson, PhD, and Robert Crawford, PhD, Armed Forces Institute of Pathology, 6825 16th Street, NW, Washington, DC 20306 After attending this presentation, attendees will have knowledge about select agent microorganisms, DNA quantitation, and microbial DNA identification techniques. This presentation will impact the forensic community and/or humanity by providing information about microbial forensic DNA identification techniques used in laboratories for the DNA identification of dangerous bacteria that are encountered in bioterrorist incidents. The microbial select agents that have been historically been weaponized in foreign nations and that pose the greatest bioterrorist threat include Bacillus anthracis spores, Yersinia pestis, Francisella tularensis, Burkholderia mallei, Brucella abortus, and Clostridium botulinum toxin, however, any microorganism or toxin can be used to create public chaos and fright in an act of biological terrorism or as a hoax. These microorganisms are cultured in a Biological Safety Level 3 (BSL3) biological containment laboratory, require special care in handling due to their pathogenicity, and are highly regulated in an effort to prevent further acts of bioterrorism. Microbial class characteristics, such as gram stain reaction, cell morphology, biochemical metabolic profiles, presence of spores and toxin production are examined in microbial forensic analyses of organisms released in a terrorist incident or hoax. Knowledge of select agent (SA) microorganisms most likely to be used as biological weapons based on historical use, recent terrorist incidents involving SA microbes, and the need for more rapid identification of these microorganisms, has prompted the quest for informative DNA identification tests. Microbial DNA identification tests require microbial strain population studies to assess the specificity, the informative value of the DNA assay for strain differentiation, and if source attribution of a strain is possible. Therefore, large repository of well characterized select agent microor* Presenting Author ganisms for the production of microbial genomic DNA for PCR assay comparison studies is being grown in the laboratory of the AFIP. Information on microbiological characteristics of the select agent genus and species, genomic information, as well as any published information is also being collected. Sensitive, specific microbial PCR techniques, performed on specimens submitted for DNA identification, are based on detection of unique genes that are present in quantified microbial DNA and can be detected whether or not the microorganism is viable. Various molecular biology techniques that have been used in the laboratory for microbial DNA identification include 16S ribosomal DNA sequencing, Amplified Fragment Length Polymorphism Polymerase Chain Reaction, Multi-locus Variable Number of Tandem Repeat Polymerase Chain Reaction, and Real-Time PCR amplification/detection of pathogen specific genes. The strategy of using combined microbiological and molecular biology techniques allows confident DNA identification, but not source attribution of select agent microorganisms. Bioterrorism, Microbiology, DNA Identification B70 An Improved Method for the Identification of Menstrual Blood Using Real Time PCR Martin Bauer, MD*, and Dieter Patzelt, MD, Institute of Legal Medicine, University of Wuerzburg, Versbacher Street 3, Wuerzburg, 97078, Germany Attendees will learn that it is possible to identify the menstrual origin of blood stains using real time PCR and that the question whether blood stains were produced by injury or by menstrual bleeding can be of crucial importance in forensic casework. This presentation will impact the forensic community and/or humanity by demonstrating a reliable method is available which allows the detection of menstrual blood markers in blood stains. Compared to a previously published paper the new technique is based on real time PCR, is much more specific and senstive and therefore suitable for the use in forensic casework. In forensic casework type and origin of stains frequently are of crucial importance. Whereas presumptive or specific tests exist e.g. for blood and semen, the question whether a stain was produced by injury or by menstrual bleeding remained unsolved over many years. Some time ago a new technique was introduced based on the detection of mRNA by reversetranscription coupled to PCR. The evaluation of potential candidate genes showed that matrix metalloproteinases were suitable markers for menstrual blood and never were positive when blood from injuries or other body fluids was examined (Martin Bauer, Dieter Patzelt. Evaluation of mRNA markers for the identification of menstrual blood. Journal of Forensic Sciences 2002; 47:1278-1283). During the last years the authors have had the opportunity to do some casework for police agencies in Germany and Great Britain. When confronted with “real” stains produced outside the laboratory it became evident that the sensitivity of the method was not sufficient because it had been only tested with artificial stains up to that time. Furthermore, an improved visualisation method with the possibility of quantifying results seemed to be helpful because it frequently was difficult to assess bands in agarose gels stained with ethidium bromide when their fluorescence was very weak. For these reasons, researcher decided to re-establish the test using real time PCR and sequence-specific probes which promised to be the perfect solution for the problems mentioned above. The use of Taqman®-probes allowed to amplify very short fragments with a size of less than 50 bp and the detction of the fluorescent dyes coupled to the probes is much more sensitive and specific than the assessment of agarose gel bands with the naked eye. Commercially available and self-designed primers and Taqman®probes for Matrix Metalloproteinases and house keeping genes were used with samples left over from previous studies and from the cases submitted 65 for examination. In addition, 60 new samples from healthy volunteers have been collected. RNA isolation and reverse transcription were performed as described previously; however, as primer for reverse transcription now random hexamer primers were used instead of an oligo-(dT)-primer. Results again confirm that the detection of mRNA specific for Matrix Metalloproteinases is a specific and sensitive indicator for the presence of menstrual blood. The use of real time PCR allowed amplification of RNA/c-DNA even in small and degraded samples using primers and probes for housekeeping genes which were negative with the conventional technique. This is particularly important because casework experience showed that it can be essential not only to prove that a given sample is menstrual blood but to state that it is not menstrual blood. With the new technique it is now possible to do this with a reasonable degree of certainty due to the sensitive and quantitative detection of housekeeping gene-mRNA. Negative result can be clearly differentiated from positive results due to the continuous monitoring of fluorescence during PCR. In this presentation the authors want to introduce the new technique and to show that now reliable and consistent results can be obtained from stains of minimal size and high degradation as it often happens in forensic practice. To demonstrate the relevance of this test casework examples will be presented in which the identification of menstrual blood played an important role in police investigations. Menstrual Blood, Real Time PCR, mRNA B71 Methodologies to Employ Porcine Tissue to Simulate Human Tissue: Determining the Efficacy of Radiation for Decontamination and the Subsequent Recovery of DNA From Tissue for PCR DNA Testing Theresa A. Caragine, PhD*, James A. Sebestyen, BS, Ewelina J. Badja, BS, Robert C. Shaler, PhD, and Mechthild Prinz, PhD, Office of the Chief Medical Examiner, Department of Forensic Biology, 520 First Avenue, New York, NY 10016 The goal of this presentation is to describe assays designed to measure DNA recovery and STR profiles from porcine tissue in order to provide a model to test decontaminated human remains. This presentation will impact the forensic community and/or humanity by presenting studies which suggest that decontamination of human tissue by irradiation is a possible solution for the safe return of reamins to families. Moreover, these methodologies for porcine tissue provide an easily accessible model for other examinations that may require human tissue samples. Currently, there are few viable alternatives regarding the identification and subsequently the interment of biologically or chemically contaminated human remains. Studies have demonstrated that irradiation of human tissue, specifically at 51 kGy, destroyed Bacilus subtilis, a surrogate for Bacillus anthracis, but preserved identifying DNA sequences. Chemical decontamination, however, will likely require much higher doses of radiation. Therefore, the laboratory of the OCME in conjunction with Titan Corporation performed a dose response with electron beam radiation and evidentiary items such as blood, semen, and saliva stains. Even at doses as high as 90 kGy, sufficient DNA to produce usable DNA profiles was recovered, although the absolute DNA yield was compromised. In order to better define the limits of DNA testing of irradiated samples, and to assess penetration through a large tissue sample, pork slabs, simulating human tissue, were irradiated at higher doses, and methodologies were adjusted to accommodate this porcine DNA. Inserting dosimeters deep within a substantial piece of pork addressed the issue of radiation penetration. With the effective dose known, one could evaluate DNA testing results taken from a pork sample proximal to a dosimeter, with certainty. Moreover, the efficacy of decontaminating tissue 66 could be studied with pork similarly “inoculated” with spore test strips for Bacilus subtilis. In this experiment, six pork shoulder shanks were sliced into four one pound slabs of boneless meat. A small portion of each of twenty slabs was sliced wherein one dosimeter and one spore strip, sealed in waterproof miniature kpac pouches, were inserted. The gap was sewn together with fishing line, and wrapped in brown bench paper. Two slabs of pork thus prepared were packaged in each of ten boxes, where one slab was placed near the bottom of the box and one near the top. Sandwiched between the two slabs were an additional dosimeter and a spore strip. Surrounding these items were dry ice and newspaper. Titan Scan, San Diego, CA, irradiated two boxes for each of the following doses: 0 kGy, 30 kGy, 60 kGy, 90 kGy, and 120 kGy. The dosimeters and the spore strips were returned to Titan Scan and Raven Laboratories, respectively, for processing. Three 25 mg samples of each of twenty slabs of pork were extracted with DNA IQ™ (Promega) according to a modified methodology developed in the laboratory on the Biomek 2000 that encompassed the use of a stronger digestion buffer and three times more resin than recommended. DNA recovery was measured with a SYBR Green I based real time PCR assay for ALU fragment. Primers described by J Walker et al. (Analytical Biochemistry, 2003) produced a 134 bp amplicon. The assay was modified for the Rotorgene 3000 and a home brew master mix with the addition of 0.5 g/ L of BSA, 8% DMSO, and 0.25 L of a 1/100 dilution of SYBR Green I (Molecular Probes). The dynamic range of the assay was from 0.78 pg to 6400 pg. The control DNA for the standard curve was procured from control slabs of pork that were extracted as described above, and measured with a spectrophotometer. Amplification of the ALU amplicon predicted successful genotyping of porcine microsatellites. Three microsatellites from the USDA database, Sw520, S0147, and TNFB, were selected due to their varying lengths, 102124 bp, 146-174 bp, and 174-213 bp, respectively. Each forward primer was labeled with 6’FAM, and collectively they were multiplexed according to the parameters described by G. Yue et al. (Electrophoresis, 1999) with additional KCL. PCR products were separated on an ABI Genetic Prism Analyzer using LIZ (ABI) as a size standard. DNA recovery and STR typing results were similar to previous experiments. Although, as radiation increased DNA recovery decreased, if sufficient DNA was present genotypes were generated. Radiation, Microsatelites, Animal Model B72 Determination of the Distribution of DNA to the Faces of Children Aged 0-5 Years Due to Normal Day-to-Day Interaction Between the Child and the Carers Victoria L. Bowyer, MSc, Eleanor A.M. Graham, MSc*, and Guy N. Rutty, MD, MBBS, Forensic Pathology Unit, University of Leicester, Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester, Leicestershire LE2 7LX, United Kingdom; and Sabine Maguire, and Beverley Ellaway, Dip Child Protection, Cardiff University, First Floor Academic Centre, Llandough Hospital, Cardiff, Wales CF64 2XX, United Kingdom After attending this presentation, attendees will learn that it is possible to obtain DNA from the skin surface of a child which can be amplified and identified and that this varies from child to child as well as at different ages. The attendee will also learn how DNA can vary in distribution across the face and neck of a child. This presentation will impact the forensic community and/or humanity by demonstrating how upon completion of phase I and phase II of this research, a series of facial maps will be produced illustrating the distribution of DNA upon the faces of children aged 0-5 years. As childcare * Presenting Author varies as the child ages these maps can be used as a template for determining how much or little DNA is transferred as the level of care changes. Variation in cases of abuse can then be investigated further with a view to determining exactly who is responsible. The goal of this presentation is to provide information on the normal distribution of DNA present on a child’s face and neck for those aged 0-5 years due to the normal day-to-day interaction between the carers and the child. Hypothesis: That it is possible to retrieve DNA from the face of a physically abused child in order to determine the perpetrator of the abuse. However, to date no one knows what the normal distribution of self and non-self DNA upon the face of children is due to the daily activities of the child for example from carer child interaction or child-child interaction. This is the first study of its type to investigate the normal deposition of DNA on the faces of children aged 0-5 years. In Britain every year three million children are victims of abuse. Many are too young or too frightened to indicate who is responsible. Victims of physical abuse are generally under the age of 5 years with the highest mortality rates being observed in those under 3 years old. Without witnesses to the event it may be impossible to identify the perpetrator and therefore prosecute them. A method of identifying the individual responsible for physically abusing the child is critically required. It is hypothesised that when an individual hits a child some of the offenders DNA will be transferred onto the child’s skin. However, before this can be confirmed one needs to know what is the normal distribution of DNA on a child. To date no information is available. The aim of this study is to provide that information for children aged 0-5 years. In order to do this the study examines the presence of DNA other than that of the child on the face, its distribution and its source. It considers in the presence of nonchild DNA whether this has arisen from the normal day to day interaction between the carer and the child. By studying children aged 0-5 years and considering the care of children and how the interaction between the carer and the child alters at each milestone it is possible to provide facial maps of the normal distribution of DNA found on the faces of children. This can then be considered if DNA is retrieved in the investigation of abuse in whether the DNA present is outside that of the normal handling pattern of the child at that age group. For the purpose of the study a facial map was designed which divided the face and neck into 12 sections. Sterile, moist swabs were used to swab each area of the face using a number of techniques. The techniques for the swabbing of children aged 0-5 years were developed such that the children could tolerate the process and yet it avoided contamination by the person undertaking the sampling. All swabs were anonymised, frozen and transported to a non-police laboratory for independent analysis. DNA was extracted from the swabs using Chelex and was quantified using Oligreen ssDNA Quantitation kit (Molecular Probes, OR, USA). DNA was amplified and analysed using AmpFlSTR SGM Plus PCR Amplification kit, ABI PRISM 377 DNA Sequencer, Genescan and Genotyper (Applied Biosystems, CA, USA). Results from phase I and phase II of a three phase study considered the facial distribution of 80 children. Each facial area showed an average quantity of DNA from the swabs less than 100ng, making accurate quantification difficult. Partial genetic profiles were obtained from the majority of the areas of the face, which were consistent with arising from the child. Extraneous alleles were observed in some areas, particularly the neck and cheeks. These partial profiles had not arisen from the child or from the person undertaking the sampling process. The results show that it is possible to obtain identifiable genetics profiles from the face using simple swabbing techniques designed within the study protocol. Facial maps for each age are being produced to show the distribution of the observed profiles. DNA Transfer, Children, Facial Mapping * Presenting Author B73 A New High Throughput Easy to Use Differential Extraction Method Allan Tereba, PhD*, Laura Flanagan, BS, Paraj Mandrekar, MS, and Ryan Olson, BS, Promega, 2800 Woods Hollow Road, Madison, WI 53711 After attending this presentation, attendees will to introduce to the forensic community a new differential extraction method that can rapidly and efficiently separate sperm and epithelial cells in a high throughput format. This presentation will impact the forensic community and/or humanity by demonstrating a new differential extraction method that will significantly reduce the amount of time needed to process a major fraction of violent crime sample types thus allowing more cases to be solved without increasing personnel. The high throughput method will allow the use of automation and will finally provide a means to significantly reduce the large backlog of sexual assault evidence. One of the main reasons for the large backlog of sexual assault samples is the difficultly in working with the evidentiary material. Typical vaginal swabs contain a mixture of victim epithelial cells in large excess over sperm cells. Unprocessed, these samples can only be analyzed using male specific markers that provide important evidence but are of limited use in searching national databases due to the inheritance and nonrecombinatorial nature of the Y chromosome. In 1985, Gill et al. developed a method to enrich for sperm cells in the presence of an excess of epithelial cells. This process relies on the fact that sperm structural proteins as opposed to epithelial cells contain a large amount of disulfide bonds that inhibit the proteolysis of these proteins. After a controlled proteolysis in the absence of a reducing agent, the sample is centrifuged in a spin basket to remove from the solid matrix intact sperm and solution containing the DNA from lysed epithelial cells. Because the resulting sperm pellet contains loose cell debris a considerable amount of contaminating solution is left and must be diluted out with serial washings and centrifugations. This process is time consuming and results in loss of sperm and variability between examiners. The authors have developed a new differential extraction method that takes advantage of the nearly two decades of experience using the standard differential extraction. After a standard Proteinase K digestion of the sample, the solid support and DNA-containing solution are centrifuged through a special material that effectively separates the sperm from soluble DNA and cell debris. The samples are washed once without centrifugation to remove any remaining soluble DNA in the sperm fraction. DNA IQ™ Lysis Buffer containing DTT is then added to the epithelial and sperm fractions. This buffer effectively lyses the sperm without need for further Proteinase K digestion. The total time for separating the sperm from epithelial cells following addition of the sample to the Proteinase K Digestion Solution is approximately 1 hour 20 minutes which includes the 1-hour Proteinase K digestion. The purification of the DNA requires 40 minutes so the total separation and purification can be accomplished in 2 hours. Because the same standard Proteinase K digestion and initial centrifugation is used to help remove the sperm from the solid support and to lyse the epithelial cells, the efficiency of these steps will be identical to what is currently available. However, only one centrifugation is required for efficient separation so the sperm recovery is better. In addition, the hands on time as well as the overall time needed to do the separation has been greatly reduced from the current method. Data will be presented on the sensitivity and successful processing of old samples. Although the new differential extraction method significantly reduces the time to process samples, there is significant time spent in transferring samples to new tubes following the digestion step. The centrifugation of samples in a single tube format also does not mesh with current automation methods. To increase the throughput of this method plasticware has been developed that allows the incubation and subsequent centrifugation of 67 samples in the same 96 deepwell format. This eliminates transfer steps where contamination can occur and makes the method fully compatible with current robotic methods. The use of this plasticware also allows the efficient extraction of reference samples on buccal swabs and blood cards to be performed with the same chemistry in the same plate as the sexual assault samples if desired. The time to process a full 96 well plate of samples using this new differential extraction method coupled with automated DNA IQ™ purification on a Biomek® 2000 workstation is approximately 4 hours of which only a small fraction of this time requires manual intervention. Differential Extraction, High Throughput, Automation B74 The Efficacy of Sample Pre-Selection in High Volume DNA Analysis Kimberley Sharpe, MSc*, Kelly Jo Walden, MSc, Roger Frappier, MSc, and Jonathan Newman, BSc, Centre of Forensic Sciences, 25 Grosvenor Street, Toronto, ON M7A 2G8, Canada The goal of this presentation is to describe the benefits of identifying acceptable sample types for analysis, with high likelihoods of success in generating crime scene DNA profiles amenable for upload to the National DNA Databank, in a high-volume casework program. This presentation will impact the forensic community and/or humanity by demonstrating the efficacy of sample selection in high volume casework and the success of obtaining DNA profiles from high-volume casework Due to caseload pressures and the volume of break and enter occurrences, these cases have had a lower priority, in the forensic community, than more serious crimes. With the advent of the National DNA Databank (NDDB) in Canada and with a recent increase in trained staff, the Centre of Forensic Sciences (CFS) developed a program for the analysis of DNA break and enter cases where no suspect has been identified. The objective was to provide information that would assist investigations of this nature, where no other avenues existed. When the program was launched in October 2001, only blood swabs were examined and only one swab per case was analysed. In August 2002, the program was expanded to include a greater range of sample types where DNA evidence stood to be recovered. This included a variety of samples such as cigarette butts, chewing gum, and swabs from discarded drinking containers. As of July 2004, approximately 2500 cases had been processed. Approximately 62% of the samples submitted were blood swabs. Of the remaining cases, a significant proportion included swabs from drinking containers (19%), cigarette butts (15%), and other miscellaneous items (4%). This review summarizes the success rate in obtaining a DNA profile from the variety of samples tested, excluding blood. The following results indicate the percentage of samples yielding a complete (AmpFlSTR® Profiler Plus™ system) STR DNA profile: cigarette butts (94%), swabs of drink containers (66%), chewing gum (100%), worn clothing with no apparent body fluid samples (32%), and miscellaneous samples such as envelopes, hair, nasal samples, and scene swabs reportedly from saliva (63%). The success of the program is exemplified by the proportion of samples yielding a profile suitable for upload to the National DNA Databank (i.e., a profile with at least 6 of 9 AMPFlSTR® Profiler Plus™ loci). The data show that on average approximately 81% of the samples accepted generated DNA profiles that were uploaded, validating the acceptance criteria, with roughly 40% of the profiles uploaded generating hits to either the Crime Scene Index or Convicted Offender Index. This casework initiative highlights the value of defining acceptance criteria based on the anticipated likelihood of success, whilst incorporating a rapid and efficient testing protocol. DNA, Casework, Review 68 B75 Automated Sample Processing and Tracking System for DNA Profiling of Single Source Samples Maja Popovic, PhD*, Elaine Schneida, BS, Siddhartha K. Sinha, BS, Anurag Bhushan, MBA, Jaiprakash G. Shewale, PhD, and Sudhir K. Sinha, PhD, ReliaGene Technologies, Inc., 5525 Mounes Street, Suite 101, New Orleans, LA 70123 The goal of this presentation is to demonstrate to the forensic community the usefulness of a tracking system and automated robotic platforms for DNA analysis of single source samples in large volume. This presentation will impact the forensic community and/or humanity by describing the usefulness of a tracking system and automated robotic platforms for DNA analysis of single source samples in large volume. The number of samples for DNA profiling has increased exponentially due to the acceptance of short tandem repeats (STRs) in forensic casework analysis and in the court system. High volume sample processing is increasing in importance in forensic laboratory, particularly for single-source samples in data base studies. The authors have designed an integrated LIMS system for tracking samples with respect to processing status, completion of the results, data acceptability, and generation of reports for ease of review. The samples are processed using Wallac DBS puncher and the extraction is performed using Qiagen BioRobot 8000DNA extraction system. Qiagen BioRobot 8000 Liquid Handling System is used for set up of fluorometric DNA quantitation, normalization of extracted DNA and PCR set up. PCR amplification is carried using 9700 GeneAmp® PCR System, fragment analysis is performed on 3100 Genetic Analyzer, and genotyping is performed with Genotyper®. All instrument platforms are compatible and interact with the LIMS software. The information required and generated at each processing station is integrated in the LIMS database. Use of robotic platforms involves minimal manual intervention and facilitates high-throughput sample processing in 96-well format. The designed system for automated sample processing and tracking minimizes the potential for error and maximizes the efficiency of DNA profiling of single source samples in high volume. Automation, DNA, Robotic B76 Strategies in Large Volume DNA Analysis of No-Suspect Casework Amrita Lal-Paterson, MSFS*, Gina Pineda, MS, Mary Burns, MS, Chris Larsen, MS, Megan Shaffer, PhD, Huma Nasir, BS, Mark Tidwell, BS, Brandi Washington, MS, Zoe Knesl, MS, Sarah Corrigan, MS, Tara Johnson, MS, Penny Reid, MS, Lesley LeBlanc, BS, Kelly Zakel, BS, Heather Overton, BS, Tabitha Benedict, MS, and Sudhir K. Sinha, PhD, ReliaGene Technologies, Inc., 5525 Mounes Street, Suite 101, New Orleans, LA 70123 The goal of this presentation is to demonstrate to the forensic community the approaches used in large volume DNA analysis of no-suspect casework. This presentation will impact the forensic community and/or humanity by describing how to process large volume DNA analysis of no-suspect casework. Analysis for short tandem repeats (STRs) in forensic casework analysis has become popular in the last decade. Further, the results from STRs have been well accepted by the court system. Because of the high power of discrimination of STRs, many States have begun examining earlier forensic case samples, including no-suspect cases for DNA analysis. The desire to process these samples for DNA evidence has overwhelmed * Presenting Author the capabilities of most laboratories. As a result, the number of backlog cases have increased in the past few years and generated a need for designing a sample processing system for high throughput. Many government forensic laboratories are now outsourcing their no-suspect rape cases to private laboratories including ReliaGene Technologies, Inc. The authors have analyzed about 1500 unscreened rape kits. The samples in these kits were screened for the presence of seminal fluid and sperm cells. Approximately 57% of the rape kits had at least one sample that was positive for either seminal fluid and/or sperm cells and 43% were negative for both seminal fluid and sperm. The cases exhibiting samples positive for seminal fluid and/or sperm cells were processed for autosomal STR analysis. In order to process such a high volume of samples, Qiagen’s EZ1 and M48 robots were integrated into the system for rapid analysis. Of the 1500 cases investigated, 50% of the cases provided complete profiles. This presentation will discuss different approaches used in the processing of samples from screening to DNA testing. Casework, DNA, No-Suspect B77 Automation of Buccal Swab Processing Cyrus K. Aidun, PhD, Georgia Institute of Technology, 500 10th Street, NW, Atlanta, GA 30332-0620; Tammy P. Northrup, JD, Darrel Oubre, Jr., BS, and Alyson Saadi, BS Louisiana State Police Crime Laboratory, DNA Unit, 376 East Airport Drive, Baton Rouge, LA 70806; Amanda C. Sozer, PhD*, DNA Technology Consulting Services, LLC, 10418 Hampton Road, Fairfax Station, VA 22039; and Pat W. Wojtkiewicz, PhD, North Louisiana Criminalistics Laboratory, 1115 Brooks Street, Shreveport, LA 71101 Attendees will learn about an automated method for introducing buccal swabs into the DNA profiling laboratory which assures sample integrity and reduces the man hours currently required to process buccal swab samples. This presentation will impact the forensic community and/or humanity by demonstrating how the current methods for introducing buccal swabs into the dna profiling laboratory are labor intensive due the need for a witness and if not performed correctly, can lead to sample mixups. The presentation will provide the forensic community with a method to rapidly and accurately introduce buccal swabs into the DNA profiling laboratory in order to efficiently and effectively address the ever-increasing number of samples which must be processed. The testing of buccal cells on cotton-tipped, Dacron®, or sponge applicators in large scale DNA profiling is desirable. The collection of the sample is non-invasive, relatively easy to perform, and collection materials are inexpensive. If collected properly, the cells on the swab yield a sufficient quantity of DNA for STR profiling. Presently, large scale sample introduction or aliquoting practices require manual cutting of the swab and placement into the correct well location in a 96 well tray or into individual tubes. Although this direct transfer of cells from the swab is more reliable and less expensive than performing an initial transfer to paper, the process is labor intensive due to witness requirements, and can lead to sample mixups, if not performed properly. With the prospect of collecting over 100,000 buccal swab samples per year in the State of Louisiana, an automated method for introducing buccal swab samples directly into the laboratory testing process was developed. This system reads and records the sample identification number, cuts the buccal swab using a non-contact laser-based method to prevent contamination and automatically places the swab into a designated location in a 96 well tray. The automated self-tracking system assures sample integrity and reduces the man hours currently required to process buccal swab samples. To demonstrate the effectiveness of this technique, cotton tipped and Dacron® buccal swabs were collected using standard methods. Two swabs were collected from each individual. One swab was cut using the noncontact cutting device, while the other was cut manually by a razor blade. * Presenting Author The DNA yields between the samples were comparable, and the samples cut with the laser showed no PCR inhibition or artifacts in the resulting DNA profiles as compared to their controls. A description of the noncontact laser cutting unit and system management software will be presented along with results from proof-of-concept studies and field experience. Buccal Swab, Automation, DNA Profiling B78 Microchip-Integrated Purification and PCR Amplification of DNA for Forensic Analysis Joan M. Bienvenue, MS*, Lindsay A. Legendre, BS, Christopher M. Easley, BS, Michael Roper, PhD, Jerome P. Ferrance, PhD, and James P. Landers, PhD, University of Virginia, Department of Chemistry, Mccormick Road, Charlottesville, VA 22904 The goal of this research project is to integrate the extraction and PCR amplification of DNA into a single microdevice capable capable of downstream STR analysis. This presentation will impact the forensic community and/or humanity by demonstrating discussing the use of next-generation microchip technology for forensic analysis of DNA. As the forensic community looks for more rapid and cost-effective alternatives to current methods, microdevices become an increasingly more viable option for improving analysis. This presentation will highlight the use of an integrated microdevice for solid-phase extraction and PCR amplification of DNA and its impact forensic genetic analysis. As genetic analysis for forensic casework continues to evolve, never before has the need for fast, accurate, analysis of samples been more pressing. Current techniques for DNA analysis require labor-intensive and time-consuming processes. These methods, though effective, have led to a dramatic backlog of casework, overwhelming crime laboratories at this time. In addition, databasing efforts are hindered by this backlog of cases. And in the current condition, many cases simply go unanalyzed. As such, research efforts in forensics have focused on improving the methods associated with the analysis of DNA to develop a more rapid and efficient assay for casework profiling. As the field looks for more rapid and cost-effective alternatives to current methods, microdevices become an increasingly more viable option for improving analysis. The application of microdevices to bioanalytical analyses has the potential to drastically reduce the time required to perform a wide variety of clinical assays. As such, microdevices are currently being designed, developed and tested to improve the efficiency of processes associated with forensic casework analysis. A fully-integrated, microchip capable of performing the steps normally carried out at the bench would not only reduce the time required to perform these tasks, but would also eliminate user intervention and potential sources of contamination, preserving more of the sample for future analysis. PCR and high-resolution DNA separations can currently be carried out on-chip, as well as solid-phase extraction (SPE) of DNA from a variety of clinical, biohazardous, and forensically significant samples. Integration of these processes is the first step towards the creation of a device with genetic profiling capabilities. The research presented here describes the chip-based approaches for executing DNA extraction (via SPE) and STR allele-specific amplification (via PCR), and how these two processes might be integrated in a single microdevice. The device and its functionality are described, along with results for extraction and amplification of human genomic DNA from sperm cells. Glass microchip devices were designed with domains specific for SPE and PCR. These were patterned using standard photolithographic techniques and the SPE domain vacuum-loaded with silica beads ‘glued’ into place with a tetraethoxysilane (TEOS) sol-gel have been shown to yield fast, efficient, solid phase extraction of DNA from a variety of biological materials. Microchip PCR amplification of forensic STR loci in sub-microliter volumes containing isolated sperm DNA is accomplished 69 using non-contact thermocycling (infrared heating and interferometric temperature sensing). Fluidic control of the movement of purified DNA, the solutions necessary for extraction, and PCR master mix into the appropriate domains on the microchip is mediated by an elastomeric valving membrane. The work reported here investigates extraction and purification and subsequent PCR amplification of DNA from semen and/or washed sperm cells on an integrated, valved microchip. This represents one of the major steps required development of a fully integrated microdevice capable of total systematic DNA analysis for forensic casework. DNA, Ccapillary Electrophoresis, Microchip B79 Joint Validation of a High Throughput Multi-Capillary Electrophoretic System Using Fluorescent Multiplex Short Tandem Repeats Bridget M. Tincher, MSFS*, Marshall University, Forensic Science Center, 1401 Forensic Science Drive, Huntington, WV 25701-3628; G. Sue Rogers, MSFS, and Angelo Della Manna, MSFS, Alabama Department of Forensic Science, 501 12th Street South, Birmingham, AL 35233; Terry Fenger, PhD, Marshall University, Forensic Science Center, 1401 Forensic Science Drive, Huntington, WV 25701-3628; and Rhonda K. Roby, MPH, Identity Quest, LLC, 13203 Astoria Hill Court, Suite J, Germantown, MD 20874 Attendees will learn procedures for high throughput processing of samples for STR analysis. The presentation will demonstrate the positive impact of collaboration with the NIJ supported Forensic Resource Network (FRN) when validating a new system. This presentation will impact the forensic community and/or humanity by validating and implementing high throughput systems in order to reduce the offender backlogs at state laboratories. Over the past eight years, state and federal laboratories have processed greater than one million convicted offender samples and subsequently uploaded the resultant data into the national level of the COmbined DNA Index System (CODIS). As of June 2004, greater than one million convicted offender samples have been uploaded into NDIS and have produced over 15,100 matches which have assisted in more than 18,100 investigations nationwide [http://www.fbi.gov/hq/lab/codis/success.htm: online]. In order for a state to upload DNA profiles developed from convicted offenders to NDIS, each sample must be comprised of the standard 13 core STR loci. Prior to this NDIS requirement, the Alabama Department of Forensic Sciences generated a state level convicted offender database containing eight (8) of the core 13 STR loci. The Forensic Resource Network (FRN) is a National Institute of Justice (NIJ) supported group of four institutions, of which Marshall University is one, that provide services, research and training to the Forensic Community. As a component of the services provided by the FRN, Marshall University Forensic Science Center (MUFSC) initiated a collaboration with the Alabama Department of Forensic Sciences (ADFS) to validate a high throughput system for short tandem repeat (STR) analysis. During the last several years, high throughput systems have evolved which increased the capacity for both processing and analyzing convicted offender samples. Previously, laboratories employed a conventional slab gel electrophoresis approach or a single capillary electrophoresis testing system. While the introduction of single capillary electrophoresis platforms required less analyst laboratory time for electrophoresis setup, the introduction of multicapillary testing platforms provided even greater throughput. Furthermore, laboratories are now using multiplexed single amplification chemistry which is able to co-amplify all of the required core CODIS loci in a single amplification, rather than the previous two (2) amplification approach which was required to obtain results of all 13 loci. 70 Once again, this advancement in forensic molecular biology techniques provides for even greater throughput. Polymerase chain reaction (PCR) multiplex amplification using commercially available kits, subsequent separation of the DNA fragments by either slab gel or capillary electrophoresis, and data analysis using specific software provides an efficient, accurate method for the compilation of DNA databases worldwide. Many laboratories have validated commercial STR kits for forensic casework, databasing applications, and parentage testing using ABI PRISM® Genetic Analyzers and/or the FMBIO® II Fluorescent Imaging Device. In order to upload data into CODIS, internal validation demonstrating reproducibility, precision, and accuracy shall be performed and documented according to the FBI Director’s, “Quality Assurance Standards for Convicted Offender DNA Databasing Laboratories” [Forensic Science Communications 2000: 2 (3): online]. Known human DNA was extracted from blood and buccal swabs. Select samples were quantitated using yield gel and/or QuantiBlot® Human DNA Quantification Kit. Extracted human cell line DNA (9947A) was included as an amplification control. Amplifications were performed in 25 L reaction volumes in MicroAmp® reaction tubes in the GeneAmp® PCR System 9700 thermal cycler (Applied Biosystems, Foster City, CA) using the Identifiler™ PCR Amplification Kit, a 5-dye multiplex kit with the necessary reagents to amplify 15 tetranucleotide short tandem repeat loci and the gender identification locus Amelogenin. The collaboration between the two agencies expedited the design, validation and implementation of the high throughput system adopted by the ADFS to increase the number of convicted offender samples analyzed in house that qualify for import into the national database. In this paper, the results of the internal validation studies performed as a result of the collaboration between the Alabama Department of Forensic Sciences and Marshall University Forensic Science Center will be presented. Supported under Award Number 2001-RC-CX-K002 from the Office of Justice Programs, National Institute of Justice, Department of Justice. Points of view in this document are those of the authors and do not necessarily represent the official position of the U.S. Department of Justice. CODIS, High Throughput, Validation B80 Streamlining the Analysis of Forensic DNA Testing: New Tools to Allow More Efficient Workflow From Processing to Final Result Cherisse Boland*, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404 Attendees will gain an understanding of the tools available for DNA testing and utilizing these tools to make decisions during the processing of biological evidence. This presentation will impact the forensic community and/or humanity by presenting an integrated system for DNA analysis. This presentation will discuss new tools available for DNA testing that allow clear decisions to be made in the initial and final stages of processing DNA samples. These decisions include: • Incorporating a screening mechanism to assess the need for further processing of DNA samples, • Detecting the presence of mixtures of male and female DNA, in order to decide a path for extraction processing and subsequent amplification, • Evaluating inhibition, • Deciding a path for extraction processing of compromised DNA samples, and • Determining if DNA results obtained from a sample meet the interpretation criteria set by the user. * Presenting Author Forensic scientists are often faced with making decisions about the best approach to take when analyzing DNA samples to obtain optimal results. However, often these decisions are made without prior knowledge of the nature of the sample. The Quantifiler™ Human DNA Quantification Kit and the Quantifiler™ Y Human Male DNA Quantification Kit can be used to assess the need for further processing of DNA samples by incorporating these assays into the workflow as a screening mechanism. The utility of the Quantifiler Human and Quantifiler Y assays in detecting mixtures of human genomic DNA from male and female sources has been demonstrated. Experiments have been designed to simulate circumstances, such as a rape case investigation, in which a small minor component of male DNA must be discerned from a high background of female DNA have resulted in successful detection of the male donor. To this end, the ability to determine if a sample contains a mixture of female and male DNA and the ability to use these assays as a quantitative interpretational aid for DNA mixtures by determining the ratio of each contributor is a new and valuable tool available to analysts. Additionally, since the Quantifiler kits allow preliminary determination of male:female mixture ratios this information will assist in selecting the appropriate method of extraction and genotyping of these samples. Further, the use of the Quantifiler kits in conjunction with the AmpFlSTR® Yfiler™ PCR Amplification Kit, produces reliable and accurate Y-haplotypes and provides the forensic scientist with a robust set of tools for Y-chromosome analysis. Part of this set of tools is a valuable haplotype population database based on analysis of the 17-locus haplotype. The Y-STR database allows the forensic scientist to estimate the frequency of a Y-STR haplotype generated with the AmpFlSTR® Yfiler™ PCR Amplification Kit. Both the Quantifiler™ Human DNA Quantification Kit and the Quantifiler™ Y Human Male DNA Quantification Kits are useful assays not only for determining the amount of amplifiable DNA, but also for evaluating the degree of inhibition in the sample. Inhibitors can interfere with the reaction at several levels, leading to different degrees of PCR efficiency and even to complete inhibition. A wide variety of PCR inhibitors have been reported, particularly in DNA samples extracted from bloodstains. Data will be shown which demonstrate the utility of the Quantifiler™ Human DNA Quantification Kit and the Quantifiler™ Y Human Male DNA Quantification Kit. Finally, until recently, genotyping projects required manual examination of all allele calls. GeneMapper® ID software minimizes this bottleneck by automating allele calling using a novel process component based quality values (PQVs) system that quickly assesses genotype quality and provides automated concordance checks. Further, for those labs that upload data to CODIS, projects can be exported into a CODIS compatible reporting format allowing easy data uploading. DNA, Quantification, Y-STR B81 Operation Iraqi Freedom: AFDIL’s Response and Challenges Faced James M. DiFrancesco, MFS*, Patricia Foley, PhD, Allison Heller, BS, Robin McDowell, MFS, Demris Lee, MSFS, David A. Boyer, MFS, James J. Canik, Brion C. Smith, DDS, DoD DNA Registry, 1413 Research Boulevard, Rockville, MD; and Lisa Pearse, MD, and Craig T. Mallak, MD, Armed Forces Medical Examiner’s System, 1413 Research Boulevard, Rockville, MD 20850 The goal of this presentation is to detail the challenges faced processing rush samples, identifying civilians, and maintaining a continuous mass fatality posture during the conflict in Iraq. This presentation will impact the forensic community and/or humanity by giving the forensic community an awareness of what goes into * Presenting Author identifying those who are lost in combat and how a lab has to adapt to challenges during a time of increased and unpredictable caseload. The opinions and assertions expressed herein are solely those of the authors and should not be construed as official or as the views of the United States Department of Defense or the United States Department of the Army. The war against Iraq began on March 20, 2003 and the combat phase officially ended on May 1, 2003, with the U.S. military gaining control over the capital city of Baghdad. Currently, hostilities continue to produce a large number of casualties. This presentation will detail the challenges faced processing rush samples, identifying civilians, and maintaining a continuous, high intensity mass fatality posture at the Armed Forces DNA Identification Laboratory during the identification process of Operation Iraqi Freedom (OIF). The Armed Forces DNA Identification Laboratory (AFDIL) supports the Armed Forces Medical Examiner System by providing DNA analysis on human remains. DNA testing may be performed as the primary source of identification; as a means to supplement other identification methods; and also for re-association of fragmented remains. Currently, one scientific method is required, although 2 or more is preferred, to identify a deceased service member. Any combination of dental records, fingerprint records and/or a DNA comparison to a known reference may be used. Reference bloodstain card samples are maintained by the Armed Forces Repository of Specimen Samples for the Identification of Remains (AFRSSIR). AFRSSIR was established in 1991 for the sole purpose of providing a direct reference sample for DNA comparisons to autopsy specimens in order that no military service member casualty should ever go unidentified. All military personnel and select civilian employees working for the military are required to have a DNA specimen on file at the AFRSSIR. The 43-day war in Iraq resulted in 142 deaths of Army, Air Force, Marine, Navy, and U.S. civilian personnel. Despite the declared end of combat operations in Iraq, casualties from the U.S. occupation in Iraq continue to be processed through the combined efforts of AFDIL, AFRSSIR and the Armed Forces Medical Examiner’s System. Currently, over 805 identifications have been made using a combination of dental, fingerprint and DNA technology. AFDIL has seen an increase in caseload, turn around time, and challenging specimens since the conflict has continued. Currently, DNA analysis is performed on all military deaths regardless of a previous identification by another scientific method. Cases in which there are no other means of identification become top priority and are completed within 72 hours from the receipt of the specimen at AFDIL. To date DNA analysis has been performed on more than 1200 samples with a 99% success rate. Deaths during Operation Iraqi Freedom (OIF) resulted from combat, homicides, accidents, suicides, and natural causes. Causes of combat deaths during OIF ranged from single gunshot wounds to massive explosions. This resulted in remains that not only varied from fully intact bodies to severely fragmented pieces, but also in the degree of decomposition. At autopsy, every suitable piece of recovered human remains was sampled for DNA testing for either identification, confirmation of identification or in the case of highly fragmented remains, re-association. The reference specimen is retrieved as required from cold storage at AFRSSIR and transported to AFDIL for analysis. A comparison is made between the DNA profiles obtained from the autopsy specimens to the DNA profile obtained from the bloodstain reference. Not all of the victims are military personnel and therefore, do not have a bloodstain reference card on file at AFRSSIR. This requires family references and personal effect to be collected and tested in order to make the identification. The personal effects that are collected are varied and have ranged from toothbrushes to laptops. Many are not suitable for testing. It is with great pride that the combined efforts of AFRSSIR, AFDIL and AFMES ensure that no service member paying the ultimate sacrifice for their country would remain unknown; but rather expeditious identification, identification confirmation and re-association will be performed to assist in laying that individual to rest. It is hoped that these efforts will provide a sense of closure to the family. Challenging Samples, Operation Iraqi Freedom, DNA 71 B82 Comparison of Buccal Cell Collection Methods Amanda M. Kleist, BS, Tammy P. Northrup, BS, JD, Darrel J. Oubre, Jr., BS, Natasha H. Poe, BS*, and Alyson Saadi, BS, Louisiana State Police Crime Laboratory, 376 East Airport Drive, Baton Rouge, LA 70806; Amanda C. Sozer, PhD, DNA Technology Consulting Services, LLC, 10418 Hampton Road, Fairfax Station, VA 22039; Mark Tourre, MSFS, Janaki Vaidyanathan, MS, MT, and Joanie B. Wilson, MT, Louisiana State Police Crime Laboratory, 376 East Airport Drive, Baton Rouge, LA 70806; and Patrick W. Wojtkiewicz, PhD, North Louisiana Criminalistics Labortory, 1115 Brooks Street, Shreveport, LA 71101 Attendees will learn the most effective and reliable buccal cell collection method(s) for DNA databasing samples. This presentation will impact the forensic community and/or humanity by presenting the most effective and reliable buccal cell collection method(s) for DNA databasing samples for a high throughput laboratory resulting from large volumes of DNA sample collections. This presentation will present the research performed at the Louisiana State Police Crime Laboratory to determine the most effective and reliable buccal cell collector samples for databasing samples. Louisiana currently has the most widespread and far reaching DNA databasing law in the country. This law expressly authorizes the collection of arrestees’ DNA for databasing purposes. The State of Louisiana has opted to utilize buccal cell collections for all arrestee testing. In order to effectuate this comprehensive databasing collection law, members of the Louisiana State Police Crime Laboratory DNA Unit along with the Unit’s contract Technical Leader and CODIS Consultant initiated research to determine the most reliable and cost efficient buccal cell collection method. The results of this research will be applied to Louisiana’s existing arrestee databasing program as well as any expansions thereto. Buccal swabs rather than blood samples are becoming the preferred method of DNA sample collection for high volume DNA profiling laboratories because of the minimized health risks, the relative ease of collection and shipping, as well as low costs associated with this method of collection. Currently, collecting an adequate buccal cell sample in a simple, fast, consistent and cost-efficient manner, while retaining the ability to store the sample, is one of the biggest challenges facing DNA profiling programs. With the anticipation of collecting over 100,000 buccal samples per year in Louisiana, a study was undertaken to compare the consistency and effectiveness of available buccal cell sampling methods and devices. The purpose of this study was to evaluate the DNA yield, the ability to obtain a successful profile, and the efficiency of sample storage using various buccal cell collection methods in order to select the most effective collection method for the State of Louisiana. Nine different collection methods were used to collect buccal cells from sixteen subjects over a five-week collection period using a collection scheme designed to minimize variability of other factors which might influence buccal cell yield. Collection methods included the Omni Swab collector, Bode Buccal DNA collector, and the Fitzco SampactTM Swab collection device. Additionally, Pur-Wraps® sterile Dacron® polyestertipped applicator, sterile foam-tipped applicator, and sterile foam-tipped applicator transferred to Whatman® Indicating FTA® Micro Card were collected. Three collections were also performed using the Pur-Wraps® sterile cotton-tipped applicator. These methods included one individual collection, one that was transferred to Whatman® FTA® Micro Card, and one that was transferred to Whatman® Indicating FTA® Micro Card. The samples were extracted according to a standard protocol which utilized the QIAamp® DNA Mini Kit (Qiagen Inc.). The amount of DNA recovered was determined by both QuantiBlot® and QuantifilerTM Human DNA Quantitation Kits (Applied Biosystems). The samples were amplified with the AmpFlSTR® Profiler PlusTM and AmpFlSTR® CofilerTM PCR Amplification Kits. Profiles were generated on the ABI Prism® 3100 Genetic Analyzer and evaluated using Genescan® analysis 72 software (version 3.7.1) and Genotyper® (version 3.7) according to laboratory protocols. A comparison of DNA yields, reliability and reproducibility of DNA profiles, quality of profiles, and ease of storage among the different collection methods will be presented. Buccal, Collection, Databasing B83 The Scientific Working Group for the Analysis of Seized Drugs (SWGDRUG) Thomas J. Janovsky, BA*, Drug Enforcement Administration, Office of Forensic Sciences, 2401 Jefferson Davis Highway, Alexandria, VA 22301; Joseph P. Bono, MA*, Drug Enforcement Administration, Office of Forensic Sciences, 2401 Jefferson Davis Highway, Alexandria, VA 22301 The objective of this presentation is to update forensic drug analysts on the Scientific Working Group for the Analysis of Seized Drugs (SWGDRUG) and on the proposals recently adopted as SWGDRUG recommendations. These recommendations include: • A Code of Professional Conduct for forensic drug analysts • Validation of Analytical Methods (with an appendix) • Minimum Recommended Standards for Sampling Seized Drugs for Qualitative Analysis This presentation will impact the forensic community and/or humanity by discussing the published recommendations which have been available since 2000 to forensic scientists around the world. These recommendations have addressed Methods of Analysis, Education and Training, and Quality Assurance issues and were developed with input from the international forensic drug analysts community. The last two sets of recommendations (Validation of Analytical Methods (with an appendix) and Sampling Seized Drugs for Qualitative Analysis) have been discussed widely in the forensic science community. During this presentation, the more significant factors from of these two sets of recommendations will be explained. Time will be allocated for questions from attendees. Representatives from the SWGDRUG Core Committee will answer questions and address the concerns of attendees. In past presentations to the American Academy of Forensic Sciences, a synopsis of SWGDRUG and the goals of the core committee have been presented. This year’s program will focus on the specifics described above. However, information is presented here for those unfamiliar with the SWGDRUG process. SWGDRUG has been in existence since 1997. The mission of SWGDRUG is to recommend minimum standards for the forensic examination of seized drugs and to seek their international acceptance. The objectives of SWGDRUG are the following: • To recommend minimum standards for forensic drug analysts’ knowledge skills and abilities • To promote professional development of forensic drug analysts • To provide a means of information exchange within the forensic drug analyst community • To promote the highest ethical standards of practitioners in all areas of forensic drug analysis • To recommend minimum standards for drug examinations and reporting • To establish quality assurance recommendations • To seek the international acceptance of SWGDRUG minimum standards The SWGDRUG core committee is comprised of representatives from federal, state and local law enforcement agencies in the United States, Canada, Great Britain, Germany, Japan, Australia, the European Network of Forensic Science Institutes (ENFSI), the United Nations Drug Control Program (UNDCP), Africa, and South America, a forensic science educator, the American Society of Crime Laboratory Directors (ASCLD), ASTM, and the National Institute of Standards and Technology (NIST). * Presenting Author All members of the core committee have worked together over the past six years to build a consensus on the development of recommendations that have impacted forensic drug analysis standards internationally. Published recommendations have been available since 2000 to forensic scientists around the world. These recommendations have addressed Methods of Analysis, Education and Training, and Quality Assurance issues. All recommendations were developed with input from the international forensic drug analysts community. Criminalistics, SWGDRUG, Drug Analysis B84 Purity Determination of Reference Drug Standards and Quantitation of Illicit Drug Samples by NMR Patrick A. Hays, BS*, U.S. Drug Enforcement Administration, 22624 Dulles Summit Court, Dulles, VA 20166 After attending this presentation, attendees will come away from the presentation with an understanding of how NMR is advantageous in the quantitation of seized drugs and determining the purity of reference standards. This presentation will impact the forensic community and/or humanity by making the attendee aware of the power of NMR as a quantitative instrument. Proton (1H) Nuclear Magnetic Resonance (NMR) Spectroscopy is a rapid, sensitive, accurate, precise, reproducible, and versatile method for determining the purity of reference standards and analyzing illicit drugs and adulterants. No reference standard of the target compound(s) is required, and aqueous and organic deuterated solvents enable great flexibility in dissolving the drug(s) of interest. For quantitative analysis, a weighed sample is dissolved in a deuterated solvent(s) with a high purity internal standard. The 7 minute NMR experiment employs 8 scans using a 45 second delay and 90o pulse. In the determination of reference standard purity, the number of quantitative values available is equal to the number of integrals of the compound. The relative standard deviation (RSD) of these signals is usually <1% for pure standards and results agree well with other instruments determining purity. Unlike separation techniques such as gas chromatography (GC), high performance liquid chromatography (HPLC), and capillary electrophoresis (CE), NMR does not require a linearity study on every compound. NMR response is proportional to the number of nuclei in a given frequency range, not on a compound’s functional groups and is not subject to stationary phase adsorption phenomena. Excellent correlation coefficients (>0.99995) were obtained using methamphetamine HCl dissolved in deuterium oxide (D2O), with maleic acid as the internal standard. Average recovery was 99.4% with RSD of 0.7%. In the analysis of seized drug samples, high spectral resolution usually results in at least one signal of the target analyte being free of interferences. In complex mixtures, multiple quantitation results of the target compound are possible in complex mixtures by subtracting the contribution of an interfering compound from an integral where the target compound also exists. This method is also applicable to the determination of multiple target components in a seized drug sample. For many exhibits good agreement was determined by NMR and other techniques. Nuclear Magnetic Resonance Spectroscopy, NMR, Quantitative * Presenting Author B85 Analysis of Seized Drug Evidence by State and Local Crime Laboratories in the United States Liqun L. Wong, MA*, and Christine Sannerud, PhD, Drug Enforcement Agency, Office of Diversion Control, 600 Army Navy Drive, E6353, Arlington, VA 22202; and Kevin J. Strom, PhD, Michael R. Baylor, PhD, Albert Bethke, PhD, and J. Valley Rachal, MS, RTI International, 3040 Cornwallis Road, PO Box 12194, Research Triangle Park, NC 27709 After attending this presentation, attendees will have an understanding of the scope and prevalence of national and regional drug evidence seized by law enforcement agencies for analysis by forensic laboratories nationwide as well as the national and regional estimates of the top 25 drugs identified and reported by the nation’s forensic laboratories from January 2004 through June 2004. NFLIS state and local forensic laboratories analyze substances secured in law enforcement operations across the country and offer a valuable and unique resource for monitoring and understanding illegal drug abuse and trafficking, including the diversion of legally manufactured drugs into illegal markets. This presentation will impact the forensic community and/or humanity by demonstrating how NFLIS is an important analytical resource for drug policy and drug control agencies to support drug scheduling efforts as well as providing timely information on drug trafficking and abuse patterns across the United State. After attending this presentation, attendees will gain an understanding of the scope and prevalence of national and regional drug evidence seized by law enforcement agencies for analysis by forensic laboratories nationwide as well as the national and regional estimates of the top 25 drugs identified and reported by the nation’s forensic laboratories from January 2004 through June 2004. This presentation will provide timely data on the variation in the distribution of controlled substances and indicators of drug availability across geographic areas based on laboratory analysis and identification from the National Forensic Laboratory Information System (NFLIS). The NFLIS program, which was established in September 1997, systematically collects results from drug analyses conducted by federal, state and local forensic laboratories. NFLIS is a database system that provides national wide drug seizure information. To date, approximately 232 individual forensic laboratories that perform drug analyses participate in NFLIS. The program’s goal is to have all 306 forensic laboratories that perform drug chemistry analyses in the United States participating within two years. Highlighted findings will include the estimated prevalence of selected “drugs of interest” and analyzed drug items by category. Quarterly Trends for the national and regional estimated number of drug items analyzed by state and local laboratories for 2001 through June 2004 will be presented. Aggregate results of drugs identified and reported by participating NFLIS laboratories representing the period January 2004 through June 2004 will be presented. The distribution of drug items by percent and number of total analyzed items in the state and local forensic laboratories will be depicted. The number and percentage of analyzed drug items for the twenty-five most frequently reported drugs, as well as the major drug categories such as narcotic analgesics, benzodiazepines, “club drugs”, stimulants, and anabolic steroids will be discussed in tables and graphs. Special study data on (1) comparison to “consumption based” data (NSDUH and DAWN); (2) drug combinations, (3) drug purity, and (4) drugs identified in strategic geographic locations as well as selected major metropolitan areas will be summarized. During the period January 2004 through June 2004, an estimated 850,000 drug items were analyzed by state and local laboratories in the United States. Cannabis/THC was the most frequently identified drug, followed by cocaine, methamphetamine, and heroin. This has been a consistent pattern for the past three years. About one percent of all reported drug items contained two or more substances, most commonly 73 heroin/cocaine. Overall, nearly 55% of drug combinations contained heroin or cocaine, or both, while approximately 20% contained methamphetamine. NFLIS state and local forensic laboratories analyze substances secured in law enforcement operations across the country and offer a valuable and unique resource for monitoring and understanding illegal drug abuse and trafficking, including the diversion of legally manufactured drugs into illegal markets. NFLIS is an important analytical resource for drug policy and drug control agencies to support drug scheduling efforts as well as providing timely information on drug trafficking and abuse patterns across the United States. Drug Analysis, Drug Seizures, Drub Database B86 Qualitative and Quantitative Values on Drug Smuggling Files Which Determined in Narcotic Laboratories, Council Legal Medicine, Ministry of Justice Turkey Salih Cengiz, PhD*, Institute of Forensic Sciences, Istanbul University, Adli Týp Enstitüsü Cerrahpasa, Istanbul, 34300, Turkey; and Tulin Cinar, MSc, Ministry of Justice, Council of Legal Medicine and Forensic Sciences, Adli Týp Kurumu, Istanbul, 34300, Turkey After attending this presentation, attendees will understand the quality and the quantity of illicit drugs seized on the routes that pass from Turkey; will understand the purities of illicit drugs in countries closer to production regions; will be briefed on the fight against drug smugling in Turkey. This presentation will impact the forensic community and/or humanity by providing valuable data about illicit drug trafficking and fighting about, so that the impact will be from police and justice departments. In this study, which was carried out at the Narcotics Division of Chemical Analysis Department, Council of the Forensic Medicine, Ministry of Justice, results of qualitative and quantitative analysis were illustrated in tables and graphs for 2,375 case reports belonging to a period of 2001 to 2003. During these three years 9,212 packages were analyzed for their contents of heroin, morphine, opium, hashish, cocaine, cannabis, cannabis- tobacco mixes, acetic anhydride, ecstasy, captagon, Legal but with red and green prescription tablets and drugs, lysergic acid diethylamide (LSD) impregnated chips, materials which were examined for their drug contents, powder and liquid amphetamines, powder methylene-dioxymethamphetamine and other solid and liquid materials which were not drugs but found in clandestine. In this study color tests, TLC and microscopic investigation were used for screening. HPLC, GC, and GC/MS were used for confirmation of the qualitative and quantitative data. It was found that among the solid materials, the highest quantity belonged to heroin and the highest quantity of tablets belonged to captagon. Ecstasy and captagon had significant increases in quantities, if all the analyzed materials are considered. The results of the quantitative analysis have shown that the purities of methylene-dioxy-methamphetamine and amphetamines still approximately at the same mean value while the purity of cocaine samples have increased about 10% in 2002 and 2003 in comparison with 2001. The purity of heroin samples have decreased about 10% in 2002 and 2003 if compared with 2001 values. In conclusion; international studies and findings concerning the production, trafficking, and purity of illegal drugs take into account the illegal production of heroin, it is believed that the traffic is shifting to neighboring countries due to 10% decrease of its purity. Illicit Drug, Composition, Trafficking 74 B87 The Application of Capillary Electrophoresis for Enantiomeric Separation of N,N-Dimethylamphetamine and its Related Analogs: Intelligence Study on Routine N,N-dimethylamphetamine Seizures Wing Sze Lee, PhD*, Man Fai Chan, MPhil, and Wai Ming Tam, PhD, Forensic Science Division, Government Laboratory, Homantin Government Offices, HKSAR, 7/F Ho Man Tin Government Offices, 88 Chung Hau Street, KLN, Hong Kong, 852, China This goal of this presentation is to demonstrate the use of capillary electrophoresis (CE) for chiral separation of the seized N,N-dimethylamphetamine (DMA) samples with aim of obtaining important information for intelligence study. This presentation will impact the forensic community and/or humanity by illustrating the significance of chiral separation of seized drugs in forensic scince so that valuable information such as intrinsic characteristics of the seized samples and clues for predicting possible synthetic methodologies can thus be obtained. Methamphetamine (MA), a powerful stimulant of the central nervous system, is one of the most popular abused drugs in Hong Kong. DMA, the N-methylated analogue of the MA, is well known of producing behavioral effects that are generally comparable to those of MA but with reduced potency. Cases related to DMA have increased in Hong Kong in the past few years. It may be due to the fact that crystalline forms of DMA and MA (‘ICE’) share similar physical appearance. Thus, DMA can be sold to abusers as MA. Most of the seized DMA samples are crystalline solids that are either in the pure form or mixed with MA. Apart from crystalline solid samples, some samples are tablets. Like ‘ecstasy’, these tablets are of different color, marking and usually mixed with a number of dangerous drugs such as 3,4-methylenedioxymethamphetamine (MDMA), 4-methylenedioxymethamphetamine (MDA), MA, and ketamine. The recent increase in the abuse of DMA in Hong Kong is of great concern to law enforcement departments. However, only a few studies on DMA have been reported in the literature compared to its analogue, MA. Therefore, a detailed study on DMA is important for deriving valuable information for drug intelligence purpose. Both MA and DMA have a chiral carbon centre at the α-position, which is known to affect their potency. For example, d-MA and d-DMA are known to have stronger stimulatory effect than their corresponding counterparts, l-MA and l-DMA. By measuring the enantiomeric excesses of the seized samples, intrinsic characteristics of these samples can be obtained. In addition to this, the enantiomeric purities of the seized drugs can provide clues for predicting possible synthetic methodologies. Identification of racemic DMA or MA indicates that these samples are derived from reductive amination of an achiral precursor phenylpropan-2-one (P2P). On the other hand, optically pure DMA or MA may have come from stereospecific reduction of the enantiopure β-hydroxyphenethylamines (i.e., ephedrine, pseudoephedrine, methylephedrine, and methylpseudoephedrine.) A number of analytical methods have been employed for the separation of chiral compounds. Both GC and HPLC are commonly used methods, however, they do have some limitations. Usually GC analysis requires tedious derivatization with chiral reagents and HPLC analysis involves the use of expensive chiral columns/mobile phase. In contrast, capillary electrophoresis is relatively simple, of low cost and has very high separation efficiency. In fact, previous study in the laboratory has successfully demonstrated the use of CE for enantiomeric separation of dl-MA and its related compounds (i.e., dl-ephedrine and dl-pseudoephedrine). In this study, an optimized CE method for simultaneous chiral separation of dlDMA, dl-MA dl-ephedrine, dl-pseudoephedrine and dl-methylephedrine is reported. The method has been subsequently applied to analyze seized DMA samples in Hong Kong. Results show that all the seized DMA samples are predominantly the d-form (>87% ee). The MA samples are of * Presenting Author various enantiomeric forms, but none of them is racemic. These results indicate that the seized samples are unlikely to be synthesized from P2P. It seems to be more likely that MA and DMA were obtained by way of reduction of ephedrine/pseudoephedrine and methylephedrine/methylpseudoephedrine respectively N,N-Dimethylamphetamine, Capillary Electrophoresis, Intelligence Study B88 Game’s Up - The Presence of Tranquilizers and Stimulants in Venison Detected by HPLC-MS Heather A. McCauley, BS*, and Rick A. Flurer, PhD, U.S. Food and Drug Administration, Forensic Chemistry Center, 6751 Steger Drive, Cincinnati, OH 45237 Attendees will learn of an unusual forensic case in which tranquilzers and stimulants were detected in processed tissue. In the absence of blood and urine, the detection of these drugs was quite challenging. The sensitivity that HPLC-MS provides was advantageous and will be discussed. This presentation will impact the forensic community and/or humanity by highlighting the advantage of HPLC-MS in the forensic laboratory setting. The sensitivity that LC-MS provides combined with the added sensitivity of the instrument in MS/MS mode permits the detection of drugs of interest in difficult matrices at low ppb range levels. The preferred matrices for detection of drug residues in biological fluids are blood and urine. While these matrices can prove to be quite challenging, in their absence, with only processed tissue available for sampling, the challenge can be amplified for the analytical chemist. This paper will discuss an unusual forensic dilemma in which the sensitivity of HPLC-MS proved to be advantageous. The sale of a specific deer for hunting is illegal in most states. Once a deer has been taken in violation of State law and transported in interstate commerce, it becomes a violation of Federal law under the U.S. Fish and Wildlife’s Lacey Act. In a recent case farm raised deer were being hunted at a private preserve in violation of State laws. In most instances, the clients traveled to the preserve from out of state, and the hides, meat, and antlers were then shipped across state lines to the hunters in their home states. The case became a joint effort between the state Department of Natural Resources and U.S. Fish and Wildlife. The Food and Drug Administration’s Forensic Chemistry Center became involved in the case when it was learned that once the deer were selected by the client, they were unlawfully tranquilized, moved to a small pen, and “reversed” (i.e., revived with stimulants) to then be killed by the client/hunter. The presence of tranquilizing and/or stimulant drugs in meat is a violation of FDA law whereby meat intended for consumption containing these compounds is considered to be adulterated. The drugs that were specifically targeted for analysis were xylazine, tiletamine, zolazepam, and tolazoline. Xylazine is an α2-adrenergic receptor agonist and acts as a sedative. Tiletamine is a dissociative agent and is pharmacologically similar to ketamine. It produces immobilization and acts as an analgesic. Zolazepam is a benzodiazepine sedative and muscle relaxant. Zolazepam also prevents seizures associated with tiletamine and is therefore marketed as the combination drug Telazol®. The drug mixture of xylazine and Telazol is commonly referred to as a “cocktail” and is considered an effective way to sedate large animals. The Telazol can be purchased as a powder and rather than reconstituting the powder with 5 mL of sterile water, 5 mL of xylazine drug solution is used and the mixture is injected intramuscularly. Tolazoline is a mixed α1 and α2 adrenergic receptor agonist and is used to reverse the effects of xylazine sedation. A number of factors made the detection of these drugs challenging for the analytical chemist. The drug dosages ranged from 250mg to 400mg * Presenting Author and the half-lives of the drugs spanned from approximately 50 minutes to 4.5 hours. However, the laboratory did not have any information regarding the time frame in which the deer were drugged before being killed. Average adult bucks range in weight from 100lbs to 250lbs, and most of the drugs readily distribute to all body tissues though tolazoline tends to concentrate in the liver and kidneys. The initial samples received in the laboratory consisted of highly processed meat. The samples were prepared using in part a method published in the “Handbook of Analytical Toxicology” which generates residues for weak acid/neutral drugs, strong acid drugs, and basic drugs. Initial sample preparation was done by weighing approximately 10g of meat and making a slurry with water. The slurry was acidified, saturated with salt, and placed in a hot water bath for 30min. The solution was then filtered and extracted with an equal volume of ethyl ether. The drugs of interest are basic and therefore only the aqueous extract was taken through further extraction procedures while the ethyl ether layer was discarded. The aqueous extract was made basic and extracted with equal volumes of chloroform. The resulting basic extract was first analyzed using GC-MS and no drugs were detected. Control deer meat was obtained to conduct spiking experiments. Based on these experiments, between 5 and 10ug/mL was the detection limit for the drugs of interest operating this instrumentation in full scan mode. Due to the sensitivity limitations and the absence of data regarding the stability of the drugs during the meat processing, detection by GC-MS did not appear to be feasible. The original sample extracts resulted in approximately 300 mL of chloroform. These extracts were taken to dryness and then reconstituted with 200µL of 10% MeOH/0.5% formic acid in water, thereby increasing the concentration by approximately 1500 fold. The samples were then analyzed using HPLCMS. The samples were analyzed with a gradient mobile phase of 0.1% formic acid and acetonitrile on a C18 column and were introduced into the MS by electrospray ionization. The sensitivity that the LC-MS provided combined with the added specificity of the instrument in MS/MS mode permitted detection of the drugs of interest in highly processed venison at low ppb range levels. Stimulants, Tranquilizers, HPLC-MS B89 The Important Role of Classical Analytical Techniques in Modern Problem Solving Approaches in Non-Routine Cases Peter R. De Forest, DCrim*, Department of Sciences, John Jay College of Criminal Justice, The City University of New York, 445 West 59th Street, New York, NY 10019 The goal of this presentation is to stimulate thinking concerning the highly varied nature of physical evidence problems that arise with regularity in criminalistics and to draw attention to the dangers of overreliance on modern technology at the expense of thoughtful scientific approaches to these problems. Modern instruments are merely very useful tools. They do not replace the experienced scientist’s brain. Introduction: Technological developments over the last few decades have made an impressive array of powerful analytical tools available to the modern forensic science laboratory. The development and refinement of chemical instrumentation over the past sixty years has revolutionized analytical chemistry and molecular biology. Physical methods have largely replaced wet chemical ones. The changes are nothing short of astounding. This has not necessarily translated into better criminalistics case solutions across the board. It certainly has improved the ability to provide better answers to relatively straightforward questions, such as “did the bloodstain found in the suspect’s car come from the homicide victim” and many similar associative evidence questions? Criminalistics is not limited to these questions. Such simple routine questions are often posed by scientifically naïve investigators or attorneys before the evidence reaches the laboratory. For many cases the laboratory-generated answers to these 75 questions suffice to provide an adequate case solution. In other case situations evidence amenable to such easily developed routine questions may not exist or such questions may not be particularly relevant in the context of the case. One of the most challenging aspects of criminalistics is that the physical evidence problems that are presented by anything but the simplest cases do not define themselves. Modern forensic science laboratories have many sophisticated tools. However, before these powerful tools can be applied to the case solution, the analytical problem must be defined. A scientific assessment of the totality of the potential evidence is required before an approach to the analysis of the physical items in the case can be designed. This is the role of the criminalist. Routine and Non-Routine Analyses: Narrow predefined questions can be answered readily with the routine application of modern instrumental techniques. For such problems, where throughput is important, they are unsurpassed. What is refered to as the microanalytical or microchemical approach is much more flexible and general. It may involve the use of many techniques including the most appropriate instrumental methods . It is an approach, not a method or technique. The experienced analyst’s brain is the most critical part of the process. The quality of the analysis does not depend primarily on instrumental parameters. It depends most critically on the experience and quality of the scientist carrying them out. With this approach the first step is most often simply looking at the sample macroscopically and microscopically. Much is learned in this way. Much is lost otherwise. Consider the information that can never be retrieved where something as simple as merely dissolving a sample in preparation for testing by certain instrumental methods comprises the first step. Such seemingly innocuous “cut and extract” or “dilute and shoot” steps often result in the loss of information including contextual information, the degree of homogeneity or heterogeneity, and something about the number and nature of physical components. For some routine analyses, where a simple question is posed, the loss of information may be of little consequence. In criminalistics applications some of these simple first steps in sample characterization may be critical. Follow-on microchemical steps may provide even richer information. A key adjective used to modify “analyses” in the above discussion is “routine.” Indeed, for routine analyses the ascendancy of instrumental methods has been an unalloyed advance. The useful role of microchemical testing in high-throughput, a routine analysis is very limited. It is with challenging non-routine problems where microchemical methods and crystal tests demonstrate their value. They are indispensable tools in a more general microanalytical approach to sample characterization. Such a general approach is needed in physical evidence investigations. Here every case is different, and a “one-size-fits-all” or “cookie-cutter” approach is inappropriate. These are the kinds of challenges that regularly arise in trace evidence problems in criminalistics and for which the microchemical approach is so ideally suited. Nothing else comes close. The microanalytical approach makes the best use of all available methods, including instrumental ones. It is this approach that defines the problem in detail and informs and guides the selection of the most appropriate tools to be employed in its solution. Crystal tests are a key component of this powerful problem solving approach. Their ready availability is essential for its successful application. The Microanalytical or Microchemical Approach: There is an unfortunate tendency among members of the public and even among some scientists to associate and perhaps even equate high tech instrumentation with science. This turns reality on its head. A minimally trained technician can often generate valid and useful data for predefined problems using modern instrumentation. This is not the case with crystal tests and microchemical approaches. In-depth knowledge of chemistry and the scientific method is necessary to exploit the microchemical approach. It takes an experienced scientist to define and redefine the problem as the analysis 76 proceeds and to select the best tools. There is no better way to deal with complex trace evidence problems. However, this approach does require experienced scientists. Unfortunately, “penny wise and pound foolish” laboratory administrators might see this as a drawback. In Summary, at first glance it might seem that many instrumental methods would offer a speed advantage over microchemical testing. However, this is often only true for routine problems where many samples are being analyzed and the instrument set-up time can be averaged over a large number of samples, so that it is a small fraction of the “per sample” analysis time. For the non-routine or “one-off” problems occurring in criminalistics, microchemical methods often enjoy a substantial speed advantage. Typically, when the need is recognized during the course of a complex analysis, the analyst can complete the test (or succession of tests) without breaking stride by having to leave the microscopy bench. T he risk of sample loss and contamination may be reduced. References De Forest, P.R., “Microchemical Crystal Tests,” letter to the editor, The Microscope, Vol. 36, No. 3 and 4, 1988, pp. 373-377. De Forest, P.R., “Recapturing the Essence of Criminalistics”, Founders Lecture, California Association of Criminalists, Science and Justice, Vol. 39, July-September, 1999, pp. 196-208. Criminalistics, Microchemistry, Crystal Tests B90 History of Microcrystal Tests for Drug Identification Hiram K. Evans, MSc*, San Bernardino County Sheriff’s Department, Scientific Investigations Division, 200 South Lena Road, San Bernardino, CA 92415-0056 Attendees will learn the extent of literature references regarding the use and validation of microcrystal tests for the identification of drugs and controlled substances. This presentation will impact the forensic community and/or humanity by supplying a histoprical perspective of use of microcrystal tests for drug identification, including a bibliography. The early history of microcrystal tests is the history of chemistry and microscopy. By the mid-1830’s toxicologists needed something besides the drastic chemical treatments applied to heavy metal poisons for application to alkaloidal poisons. While any history moves forward in small steps, microcrystal tests in forensic science have a series of watershed dates. 1865 brought Helwig’s Das Mikroskop in Der Toxicologie and Wormley’s Microchemistry of Poisons. By 1921 and the publication of Behrens-Kley’s Organische Mikrochemische Analyse and Stephenson’s Some Microchemical Tests for Alkaloids forensic science had expanded to include the identification of controlled drugs. 1934 and 1935 saw the publication, respectively, of Amelink’s Schema zur Mikrochemischen Identifikation von Alkaloiden and Rosenthaler’s Toxicologische Mikroanalyse. From the 1920’s through the 1960’s, frequent collaborative work was performed and published in JOAC, expanding application and introducing acid reagent media. 1969 was probably the greatest year with publication of E. G. C. Clarke’s Isolation and Identification of Drugs and Charles C. Fulton’s Modern Microcrystal Tests for Drugs. Publications on microcrystal tests have decreased in number, concentrating on determination of isomeric forms, but the tests remain part of some training programs, are included in new ASTM Standard Guides, and are accepted by ASCLD/LAB for use in accredited laboratories. Microcrystal Tests, Drugs, Identification * Presenting Author B91 Practical Identity Using Microcrystal Tests B92 Wayne Moorehead, MS*, Orange County Sheriff-Coroner, 320 North Flower Street, Santa Ana, CA 92703 After attending this presentation, attendees will learn the role of microcrystal tests in analytical schemes and their usefulness in criminalistics. This presentation will impact the forensic community and/or humanity by reviewing of some aspects of the essence of criminalistics and suggestions for increasing the visibility of this useful method of analysis. Most forensic scientists with a modicum of interest in forensic science when asked for the definition of their profession will be able to state that it is the application of science toward civil and criminal law matters. Ask them to describe the core principles of criminalistics and, for many, silence results. Near the hub of the profession of criminalistics resides the practical identity of evidence. To the criminalist identity has two meanings – to make an identification and to make an individualization. The evidence determines which of the two or if both concepts are to be pursued. Criminalistics is the science of individualization – the pointing to or identity of one object or person to the exclusion of all other objects or people. Often the criminalist must identify the evidence before individualizing it to a unique person, place, or thing. In criminalistics, making an identification of an unknown substance is not unlike other sciences. When the criminalist has an unknown powder suspected of being a drug, an identification of the drug is made by matching the properties of the questioned drug to a known drug having the same properties. Microcrystal tests have been a part of the practical identity of routine controlled substance analyses for many decades as well as for unknown substances in trace evidence analysis. There are two types of science in practice. The first type is theoretical. The development of new materials and technology requires determining structure and properties, and exemplifies what the public perceives as science. This science identifies never before identified chemicals through structural elucidation by using an array of instrumentation and methods. This type of identification, that of identifying a “true unknown” with a variety of instrumental methods, never occurs in forensic science. The second type of science involves the practical identification or comparative analysis of materials. A comparison of the unknown against a database of known standards is conducted. The standards may be in the form of spectra or the results of chemical reactions called microcrystal tests. The comparison of a microcrystal test result from a questioned sample to a known drug standard is equivalent to the common practice of comparing the infrared or mass spectrum of a questioned sample to a known drug standard. The inclusion of microcrystal tests as the confirmatory tests of an analytical scheme for common controlled substances results in the practical identity of the drug, providing a savings of time per case, and greater case output with no reduction in the quality of the analyses. All scientists, whether from outside the field of criminalistics or from within, must understand the role of microcrystal tests for common drugs of abuse: that of being the last part of a scheme of analysis. For unknowns in trace evidence, the microcrystal tests are used in conjunction with optical properties or other microscopical or instrumental methods. In order to improve the visibility of microcrystal tests some suggestions are offered. 1) Give presentation on microcrystal tests at large venue scientific meetings 2) Publish papers in widely read and respected scientific journals 3) Respond to misleading and misinformed articles on color spot tests and microcrystal tests 4) Do not use an article as a reference that is misleading or misinformed 5) Emphasize the benefit of microcrystal tests for teaching qualitative analysis in inorganic and organic chemistry – e.g., limited amounts of material and safety to students 6) Encourage other scientists to use microscopy and microcrystal tests 7) Join scientific organizations to provide correct information on the use, value, and limitations of microcrystal tests. Drugs, Microcrystal Test, Identification * Presenting Author SWGDRUG, ASTM, and SDSO: How the San Diego Sheriff’s Lab Uses Microcrystalline Tests in Drug Analysis Marty C. Fink, BA*, San Diego Sheriff’s Crime Laboratory, 5255 Mount Etna Drive, San Diego, CA 92117-6912 Attendees will learn that there is still a place for 19th century chemistry using microcrystal tests in a 21st century laboratory. This presentation will impact the forensic community and/or humanity by showing that the use of microcrystal tests still has a place in a modern crime laboratory. Microcrystalline tests are old; they’re anachronistic; they are good for screening only; they offer no structural elucidation; they produce no reviewable data. All of these arguments have been used to preclude the use of microcrystalline tests in drug analysis. However, 19th century chemistry still works and has a place in a 21st century lab. The Scientific Working Group for the analysis of Seized Drugs (SWGDRUG) formally adopted a set of recommendations on Education and Training, Quality Assurance, and Methods of Analysis of seized drugs in May, 2000. In the forward, SWGDRUG “strongly urges the adoption of this recommendation by any laboratory involved in the analysis of seized drugs.” The original recommendations excluded any use of microcrystalline tests as part of the Minimum Standards for Forensic Drug Identification. Several letters and discussions later, microcrystalline tests were included in Category C, the least discriminatory of the categories along with color tests, melting point, and UV spectrophotometry. More letters and discussion moved them to Category B where they currently reside. The SWGDRUG recommendations were initially submitted to ASTM prior to the 2002 ASTM meeting in Atlanta for adoption as standard methods. They were defeated and returned to committee for revision. At the February 2004 meeting in Dallas, after a few modifications, ASTM officially adopted the SWGDRUG recommendations as the standard for drug analysis. In the SWGDRUG identification scheme, if a Category A method is not used, two uncorrelated methods from Category B must be used. The San Diego Sheriff’s Lab uses a combination of color screening tests, microcrystalline tests, and thin layer chromatography for the majority of the powder identifications. Each analyst receives training in basic microscopy and a core training module in the use of microcrystalline techniques as part of the training program. The examiner is ultimately given 25-30 qualifying samples in which only color spot tests, TLC, microcrystalline tests, and FTIR can be used. GCMS training is a separate module commencing only after the drug chemist has mastered the above four techniques. A second set of qualifying samples is then given in which GCMS may be used. Microcrystal Tests, SWGDRUG, ASTM B93 The Characterization of Reloading Smokeless Powders Toward Brand Identification Wayne Moorehead, MS*, and Annie Tibbetts, BS, Orange County SheriffCoroner, 320 North Flower Street, Santa Ana, CA 92703 After attending this presentation, attendees will have an understanding of the value of morphology, micrometry, and instrumental methods in the brand identification of smokeless powders. This presentation will impact the forensic community and/or humanity with the use of the database of information by providing investigators and prosecuting attorneys with the brand or a short list of brands of smokeless powder used by a bomber. Additionally, the database may be available on CD at a later time for use by other laboratories. 77 On completion of the presentation, the listener will have an understanding of the value of morphology, micrometry, and instrumental methods in the brand identification of smokeless powders. By using morphology, micrometry, and other methods, unique brand identification or a short list of possible smokeless powders from an unknown powder is possible. A sample size of 148 smokeless powder brands, with a few repeats, was examined for potential identifying features such as morphology, color, texture, coating, dimensional measurement, weight, and chemical content. The questioned powder sample must contain a sufficient number of unburned powder kernels for comparison. While the government closely monitors explosives, canisters of smokeless powders can be purchased over-the-counter by sport shooters and hunters for reloading ammunition or by bombers for improvised explosive device construction. Reloading or canister smokeless powders offer an available relatively inexpensive explosive for the criminal element. Explosive fillers from pipe bombs submitted to crime laboratories frequently contain smokeless powder. Providing smokeless powder brand identification information to an investigator can assist in the investigation or trial of a bombing suspect. Smokeless powders were subjected to different methods of analysis to build a library database for future comparisons for brand identification or provide a selective list of possible powders. Macroscopic and microscopic features were noted. Unless a unique identifier, such as a colored ‘dot’, was present the macro-scale examination could not produce a single, unique brand. Using a stereomicroscope with a digital camera and semi-automated measurement software, at least 25 random kernels (and up to 100 random kernels) from each of the 148 samples were measured for their relevant dimensions. Use of micro-morphology and other microscopical features to categorize the powders eliminated significant non-conforming brands for further comparison. Micrometry measurements were statistically evaluated against like morphology. The dimensional parameters were treated to Bonferoni-Dunn statistical analysis. The standard T-test was rejected because it is used for one-against-one comparisons while Bonferoni-Dunn provides one against many, taking into account the effect of multiple comparisons. Often characterization of features such as micro-morphology, color, texture, and coating, along with dimensional measurements permitted unique or limited number of possible smokeless powder brands. The mass of sample of the various powder brands using triplicate samples of 50 kernels at a time for measurement further characterized the powders. Gas chromatography with mass spectrometry was used to characterize the smokeless powders with positive results. Of the 148 samples characterized, most brands could be identified uniquely while some samples resulted in a short list of possible brands. Chemical content was able to resolve several of these. Smokeless Powder, Morphology, Micrometry B94 The Development of a Comprehensive Scheme for the Analysis of Electrical Tape Using Instrumental and Chemometric Methods Amanda B. Sturdevant, BS*, School of Criminal Justice, Michigan State University, East Lansing, MI 48824; and Leanora Brun-Conti, BS, Edward C. Bender, BS, and John V. Goodpaster, PhD, Forensic Science Laboratory, Bureau of Alcohol, Tobacco, Firearms and Explosives, 6000 Ammendale Road, Ammendale, MD 20705 After attending this presentation, attendees will gain an understanding of the capabilities and limitations of some of the techniques available for the forensic analysis of electrical tape. Furthermore, this research will provide the forensic science community with an effective analytical scheme for examining and comparing questioned and known samples of electrical tape. 78 Polyvinyl chloride (PVC) tape, more commonly known as electrical tape, is often encountered during explosive investigations. Electrical tape may be used in the construction of improvised explosive devices (IEDs) for wire insulation, for sealing openings, and for securing fuses and other components onto a device. If tape is recovered and submitted as evidence, an analyst may be asked to compare the questioned tape with a known source, often a roll of tape obtained from a suspect. On rare occasions a physical match of the tape ends can positively identify the questioned tape fragment as having originated from the known source. However, when such individual characteristics are not present, the analyst must resort to comparisons of class characteristics for association or elimination. Most often these include physical dimensions, surface texture, and chemical composition. Before carrying out any research study of the class characteristics of electrical tape, a number of challenges must be overcome. First, a detailed understanding of the inherent heterogeneity of the sample population, arising from both chemical composition and product distribution, is needed. Only then can a representative sample of the population be obtained. However, most formulations of electrical tape are proprietary and the manufacturers will not disclose the identity of some of the raw materials used. This limits what information is known about the composition of the product. With regard to distribution, tape from the same manufacturer may often be marketed under many different brand names. Chemically and physically the tapes are considered to be the same, while commercially they are different. Next, following sampling, the use of multiple analytical methods is necessary in order to fairly evaluate the relative merits of each individual method. Finally, using quantitative methods of analyzing the results allows for more reliable conclusions. There have been numerous articles published regarding the forensic analysis of adhesives and a few that focus specifically on electrical tape; however, none of these have directly addressed the above-mentioned issues. In this study, discussions and plant tours with tape manufacturers and careful attention to manufacturing/product codes has allowed for a greater understanding of the tape industry. The sampling methodology involved acquisition of known exemplars directly from a manufacturer as well as the traditional route of purchasing tape from commercial sources, but with a better understanding of the true manufacturer of a sample. Specifically, nine rolls of tape, including three rolls from three different batches of three different types of tape, were obtained directly from a manufacturer, 3M. Also, 32 other samples from 18 other brands representing a total of 10 manufacturers were obtained. All the samples were analyzed by the following methods: Attenuated Total Reflectance (ATR) Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy with Energy Dispersive Spectrometry (SEM-EDS), Pyrolysis-Gas Chromatography-Mass Spectrometry (Py-GC-MS), and High Temperature GC-MS. The results from each of the techniques were evaluated individually and in various combinations, via principal components analysis (PCA) to determine their relative ability to distinguish between the rolls of tape. With this information, an analytical scheme for electrical tape was developed. Preliminary results demonstrated that between the three brands of 3M tape obtained directly from the manufacturer, elemental composition alone can discriminate between the general-purpose brand and the professionalgrade brands. Initial experiments with chemical extraction of the plasticizer have shown that High Temperature GC-MS may be able to elucidate more information about this particular component of electrical tape than traditional methods have previously allowed (e.g. FTIR and Py-GC-MS). Also in this study, FTIR methods examined in previous work were employed to screen the samples and subsequent separations/extractions provided further discrimination. Electrical (PVC) Tape, Improvised Explosive Devices (IEDs), Chemometrics * Presenting Author B95 Quantitation of Male/Female Mixed DNA Samples Using the ABI PRISM® 7000 Sequence Detection System Ashley J. Hinkle, BS*, Marshall University Forensic Science Graduate Program, 1401 Forensic Science Drive, Huntington, WV 25701 Attendees will learn how the ABI PRISM® 7000 Sequence Detection System, in conjunction with the Quantifiler™ Human and Quantifiler™ Y Human Male DNA Quantification Kits, can be used to determine DNA concentration in male/female mixed source samples. This presentation will impact the forensic community and/or humanity by describing a relatively new technology that can be implemented in a forensics laboratory. Mixed Male/Female DNA samples are often encountered in the forensic laboratory. Quantitation of each DNA source in a sample can be an important tool in interpreting the final STR profile. The Quantifiler™ Human and Quantifiler™ Y Human Male DNA Quantification kits, in conjunction with the ABI PRISM® 7000 Sequence Detection System use real-time PCR to amplify human and human-male DNA, respectively. Male/female DNA mixtures of known ratios will be amplified with both kits to determine the concentrations of total human and total male DNA present in each sample. Theoretically, the difference between total human DNA and total male DNA should equal the amount of total female DNA in each sample. This project will determine if the ABI system can accurately predict the amount of male/female DNA in a mixture. Thirty-eight mixtures of male/female DNA were quantitated with both the Quantifiler™ Human DNA and Quantifiler™ Y Human Male DNA quantification kits developed by Applied Biosystems. When the female contribution in a male/female mixture was the minor component and the total DNA concentration was greater than approximately 1.48 ng/ l, the female DNA concentration could not be accurately determined by subtracting the total male DNA concentration from the total human DNA concentration. When the male contribution in a male/female mixture was the minor component, the presence of an excess contribution of female DNA did not interfere with the performance of the Quantifiler™ Y Human Male DNA quantification assay. The Quantifiler™ Y Human Male DNA Quantification kit did not produce linear results at higher DNA concentrations (greater than 20 ng/ l). This non-linearity was reproducible, but varied in severity from assay to assay. In addition, it cannot be understated how essential correct pipetting techniques and properly calibrated pipettes are to accuracy and precision. Small errors in pipetting can translate to substantial deviation in the final quantitation results. Quantifiler, Quantification, DNA B96 Reducing Stutter Artifacts in Forensic DNA Analysis Using Polyhydric Compounds Elisabeth Schoneau, BS, Lindsey Wander, BS, and Sulekha R. Coticone, PhD*, California State University, Fresno, 2555 East San Ramon Avenue, Fresno, CA 93740 Attendees will learn improvements in the analysis of short tandem repeats in DNA typing. This presentation will impact the forensic community and/or humanity by improve methods in analysis of DNA typing. Current methods for PCR amplification of short tandem repeat (STR) loci result in an artifact named “Stutter.” These stutter products are shorter than the target allele by multiples of the repeat unit. Due to the presence of the stutter artifact, multipeak patterns are obtained with STRs which results in complication in the interpretation of results specifically in mixture analysis in forensic samples and measuring microsatellite instability in cancer diagnosis. * Presenting Author Polyhydric compounds and specifically polyols have been shown to increase the thermal stability of proteins under stressful conditions. Additionally polyols have been shown to interact with the polynucleotide solvation sites replacing water surrounding the double helix. Due to the combination of the functions of polyols mentioned above, the processivity of the enzyme may be enhanced by the presence of the polyols during the amplification of short tandem repeats. A screening method has been developed to measure the effect of polyhydric compounds on reducing the stutter artifact. The screening method involves the use of a PCR fluorescent multiplex system specifically used for detecting DNA in Forensics. The multiplex system (Profiler Plus ID) is combined with the use of various polyhydric compounds. Following PCR using the fluorescent multiplex the products are analyzed on a capillary electrophoresis system (ABI Prism 310) and detected using the Gene Mapper ID software system. The results are determined based on stutter percentage ratios. The stutter percentage ratio is defined as the ratio of the stutter allele to the main allele peak. Among the polyhydric compounds tested, there was a 2-20% decrease in stutter as compared to the control in all loci. Arabinose, sorbitol, glucose and sucrose contributed to the maximum decrease in stutter reduction. Further studies include varying the concentration of the polyhydric compounds and testing osmolytes for further reduction in the stutter artifact. This will improve genotyping and allele assignment of microsatellites by eliminating errors in genotyping thereby improving the sensitivity of the technique. In the forensic field specifically samples containing mixtures of DNA samples will be easier to decipher simplifying analysis. STRs, Stutter, Polyols B97 Using Microscopy to Predict Success or Failure of a Differential DNA Extraction Julie E. Golden, MS*, Jefferson Parish Coroner’s Office, 2018 8th Street, Harvey, LA 70058 Attendees will learn the results of a study examining the usefulness of microscopy in order to predict the success or failure of a differential DNA extraction. This presentation will impact the forensic community and/or humanity by demonstrating that microscopic evaluation of sexual assault evidence is a valuable tool for determining whether or not the traditional differential DNA extraction method will be effective. Differential DNA extraction is a commonly used method of examining sexual assault evidence in the forensic DNA laboratory. This extraction method allows the DNA analyst to chemically separate sperm donor DNA from epithelial cell donor DNA resulting in two fractions. A differential extraction failure is defined as incomplete removal of the epithelial cell portion of the sample from the sperm cell fraction resulting in a DNA mixture of the two donors or the presence of the epithelial cell donor only in the sperm fraction. A sperm cell fraction that results in a mixture of epithelial and sperm donor DNA can be problematic when performing statistical analysis of the sample; decreasing the weight of the DNA test results in court. In addition, sperm cell fractions resulting in mixtures can make productive searches of DNA databases, such as CODIS, more difficult. Microscopic slides have been prepared from 325 vaginal swabs. These slides have been stained via the traditional Christmas tree staining procedure and examined microscopically. The number of epithelial cells and the number of sperm cells was rated for each slide. In addition, DNA from the vaginal swabs has been extracted using a traditional organic differential procedure. The sperm cell fractions of these extractions have been amplified using the AmpFISTR Profiler Plus and COfiler amplification kits and analyzed via the ABI Prism 310 Genetic Analyzer. The resulting sperm fraction DNA profiles were examined in conjunction with the microscopic data in order to determine the correlation, if any, between the microscopic data and the mixed or single source nature of the DNA profile results. 79 The findings of this study show that microscopic ratings indicating a large number of epithelial cells regardless of the number of sperm cells cannot predict a failure of the standard differential DNA extraction procedure. Likewise, microscopic ratings indicating a small number of sperm cells regardless of the number of epithelial cells also cannot predict a failure of the differential DNA extraction procedure. However, samples that contain both a small number of sperm cells and a large number of epithelial cells will result in a failure of the differential DNA extraction procedure in 70% of cases. A modification of the standard organic differential method was utilized on a subset of 20 samples that originally resulted in a sperm fraction DNA mixture. However, adding an additional aliquot of Proteinase K during the epithelial cell digest and increasing the number of sperm cell washes only improved samples with microscopic ratings indicating larger numbers of sperm present. This modification did not increase the likelihood of obtaining a single source sperm fraction for samples with the characteristic microscopy (low sperm/high epithelial) indicating a mixed DNA profile is likely. Therefore, these results indicate that perhaps epithelial cell DNA is bound somehow to the sperm heads or is in some other way difficult to remove from the sperm fraction via the traditional procedures. This study has demonstrated that microscopic evaluation of sexual assault evidence is a valuable tool for determining whether or not the traditional differential DNA extraction method will be effective. If an effective alternative differential DNA extraction method was available, samples identified via microscopic evaluation likely to result in sperm cell fraction mixtures using the traditional organic method could be diverted, thereby increasing the number of single source sperm fraction profiles. DNA, Mixture, Microscopy B98 Development of a Semi-Automated Robotic Platform for the Extraction of Forensic Evidentiary Samples Using the Tecan Genesis 150 David E. Sipes, MS*, Jason E Kokoszka, PhD, Jonathan Freedman, BS, and Robin W Cotton, PhD, Orchid Cellmark, 20271 Goldenrod Lane, Germantown, MD 20876 Attendees will learn a cost-effective, semi-automated robotic extraction platform designed to improve throughput and reduce the labor costs commonly associated with manual extraction procedures. This presentation will impact the forensic community and/or humanity by providing the forensic community with a method by which to implement a robotic extraction platform. With the vast number of backlogged evidence samples and the infusion of NIJ funding with which to process these samples, the forensic DNA community has been challenged to develop a higher throughput approach to sample processing in response to a growing demand for its services. While the utility of robotic platforms has been clearly demonstrated in clinical laboratories and more recently in the extraction of forensic database samples, automated DNA extraction techniques have now broadened in scope to include forensic evidence samples. Robotic alternatives to the conventional manual techniques must be designed such that the automated systems are as adept at handling the diverse variety of forensic samples as their manual extraction counterparts. Previous validation studies in the laboratory and others have demonstrated that the Qiagen DNA extraction kit is a suitable alternative to the organic method of DNA extraction. It was the objective of this validation to develop a robotic alternative to the liquid handling steps in the Qiagen DNA extraction protocol. After the manual addition of lysis buffers to the sample and the subsequent incubation, the lysates are then transferred to the Tecan Genesis 150 for the remainder of the Qiagen procedure. When the extraction is 80 complete, the samples are eluted in a microtiter plate and an aliquot of the extracted DNA is robotically transferred to a second plate for quantitation. Because the customary elution volume from the Qiagen column is 200ul, evidence samples are subsequently concentrated using a plate-based microcon. Standard samples usually contain sufficient quantities of DNA and no further concentration steps are necessary. All of the liquid transfer steps in the Qiagen DNA extraction protocol have fixed volumes with no variation from sample to sample. When the volumes vary (i.e., quantitation setup), a worklist containing sample specific volumes and well locations is generated by an internal LIMS system and is exported to the Gemini (Tecan) software. Ensuring that the robotic sample handling schematic is free of contamination is at the forefront of the developmental process. Once the liquid transfer steps were defined and the instrument performance files were optimized, the deck layout was designed to facilitate the most efficient movement of the robotic arm as well as to minimize the occurrence of tips crossing over open sample tubes and reagent troughs. Because of the wide variety of DNA amounts encountered in evidence samples, a comprehensive contamination study was performed to ensure that any occurrences of contamination would be detected. A series of runs using highly concentrated DNA samples arranged in a checkerboard pattern were performed to detect contamination that may have occurred during robotic processing. To evaluate the efficiency of the robotic versus manual extraction method, lysates from individual samples were evenly divided into two tubes. One tube was extracted manually and the other robotically. A quantitative comparison of DNA recovery showed no significant differences in recovery between the robotically extracted samples and those that were extracted manually. While the immediate benefits of extraction automation can be realized in terms of reduced labor cost and higher throughput, the downstream advantages are often overlooked. By eluting samples in a microtiter plate format, subsequent plate-based sample transfers (i.e., amplification and/or instrument sample preparation) are more readily performed using a multichannel pipette, improving efficiency and precluding the need to label tubes. This poster will show that the extraction efficiency of the Qiagen kit coupled with the versatility of the Tecan Genesis Liquid Handling robot makes the extraction of standard and evidence samples very amenable to automation. Additional automation considerations including low volume pipetting, optimization of liquid classes and the judicious choice of deck layout will be presented. Tecan Genesis, Qiagen, Automated DNA Extraction B99 A Comparison of Y-STR Multiplexes Cassie L. Johnson, MS*, Orchid Cellmark, 2600 Stemmons Freeway, Suite 133, Dallas, TX 75207; Ryan Reynolds, BS, UNTHSC, 3500 Camp Bowie Boulevard, Fort Worth, TX 76107; and Rick W. Staub, PhD, Orchid Cellmark, 2600 Stemmons Freeway, Suite 133, Dallas, TX 75207 Attendees will learn how various Y-STR multiplexes compare to one another and their applicability to forensic casework. This presentation will impact the forensic community and/or humanity by providing an understanding the available types of Y-STR kits and their advantages/disadvantages will enable crime laboratories to make an informed decision when selecting a Y-STR multiplex. Y-STR testing enables forensic DNA analysts to examine the male contribution of an evidentiary sample without interference from female DNA. This testing can be quite valuable for cases in which traditional autosomal STR testing has been unsuccessful or has yielded inconclusive results. The proportion of forensic cases utilizing Y-STRs has increased as DNA analysts, law enforcement agencies, and lawyers learn of the advantages of Y-STR testing. In response to the increased demand for Y-STR analysis, several multiplexes have been developed to allow for the simultaneous amplification of Y-chromosome loci. The present study compares a * Presenting Author Y-STR 10-plex already employed in casework at Orchid Cellmark, as well as two commercially available multiplexes (Applied Biosystems’ Y-Filer and Promega’s PowerPlex®-Y). The sensitivity and reproducibility of each multiplex, as well as analysis of female:male and male:male mixtures has been examined. The advantages and disadvantages of each multiplex as they relate to its applicability to forensic casework will also be presented. Y-STR, Multiplex, DNA B100 Evaluation of Poly(ethylene oxide) as a High-Speed, High-Resolution Sieving Matrix in Shortened Capillaries or Microchips for Forensic STR Analysis Joan M. Bienvenue, MS*, Kate Wilson, BS, Jerome P. Ferrance, PhD, and James P. Landers, PhD, University Of Virginia, Department Of Chemistry, Mccormick Road, Charlottesville, Va 22904 The goal of this research project is to demonstrate the use of poly(ethylene oxide) (peo) as a high-resolution sieving matrix for forensic analysis of DNA. This presentation will impact the forensic community and/or humanity by discussing the evaluation of an alternative polymeric sieving matrix for high-resolution separation of DNA, as well as its potential inclusion in next-generation microchip technology for DNA analysis. Capillary electrophoresis is now the sanctioned method for separation and analysis of short tandem repeat (STR) fragments of DNA for forensic application and genetic mapping. Increased speed, separation efficiency, and automation have made the technique indispensable in the forensic laboratory. In order to accomplish the high-resolution separations required for STR analysis, a limited number of polymeric solutions have been evaluated systematically for suitability in this application. Linear polyacrylamides have become the separation matrix of choice because of their stability, reliability and ability to yield high-resolution separation DNA fragments differing in length by a single base. However, the high cost of commercial products, poor resolution over shorter capillary distances resulting in longer separation times, and the high viscosity of some polymers, has created the need to search for alternative sieving matrices. Though current commercial products accurately and reproducibly separate fragments with high resolution, in lengthy capillary systems, their cost and separation efficiency over short distances become prohibitive when making the shift from capillary-based separations to microchip-based systems. These considerations become increasingly significant when attempting to translate conditions from capillary-based methods to the microchip platform, where channel length and total device area of are utmost importance. As the forensic community moves towards fully integrated micro devices capable of sample preparation and full genetic analysis, choice of separation matrix will play a major role in device design, analysis time, and ease of use. Thus, as polymers are evaluated using capillary electrophoresis, attention must be paid to the feasibility of the sieving matrix for microchip-based analysis. Critical evaluation of polymer viscosity, resolution over short distances, device preparation, reproducibility of separations, ease of polymer preparation, and cost of analysis must occur when developing a polymer for separation of STR fragments for forensic genetic analysis with the goal of translation to the micro device. The work presented here describes the evaluation of poly(ethylene oxide) (PEO), an inexpensive, commercially-available polymer, for separation of STR fragments in capillaries and potentially for inclusion in the microchip platform. Accurate, repeatable separations of STR fragments with single base pair differences are reported using a denaturing, lowviscosity, low-molecular weight solution of PEO. The effects of buffer composition, capillary temperature, polymer weight and concentration and capillary length are described. A variety of polymer weights are explored at varying concentrations to determine the polymeric conditions that result * Presenting Author in the highest resolution separation. Shortened capillaries were used to simulate separation lengths similar to feasible microchannel lengths in analytical microdevices for evaluation of polymer properties. Reproducibility of the separation is evaluated, as well as the use of epoxypoly(dimethylacrylamide)(EPDMA)-coated capillaries for minimization of electroosmotic flow (EOF). Urea concentration and capillary temperature are also evaluated and optimized conditions for high-resolution separation of samples in shortened channels are presented. Comparisons of PEO to conventional commercial products are highlighted and the appropriateness of this sieving matrix for translation to the microdevice is appraised. DNA, Capillary Electrophoresis, Microchip B101 Mexican (Chiuahua) Population Data for the 15 STRs Loci Included in the Identifiler Kit Luis J. Martinez-Gonzalez, MS, Laboratory of Genetic Identification, Department of Legal Medicine, University of Granada, Granada, GR 18012, Spain; M. Moguel, MS, State of Chihuahua DNA Laboratory, D.G. Servicios Periciales, Chihuahua, Mexico; E. Martinez-Espin, MS, Laboratory of Genetic Identification, Department of Legal Medicine, University of Granada, Granada, 18012, Spain; C. Entrala, PhD, J.C. Alvarez, MS, and F. Fernandez-Rosado, DDS, Laboratory of Genetic Identification, Department of Legal Medicine, University of Granada, Granada, 18012, Spain; José A. Lorente, MD, PhD*, University of Granada, Department of Legal Medicine, Av. Madrid, 12, Granada, 18012, Spain; Miguel Lorente, PhD, and E. Villanueva, MD, Laboratory of Genetic Identification, Department of Legal Medicine, University of Granada, Granada, 18012, Spain; and Bruce Budowle, PhD, FBI, FBI Academy, Quantico, VA 22135 The goal of this presentation is to present the results and parameters of forensic interest (HWE, PD, PE) of the Mexican population for the 15 STR loci included in the Applied Biosystems Identifiler kit. This presentation will impact the forensic community and/or humanity by demonstrating data of great interest for the forensic community, since they are necessary to perform statistical calculations, both in the paternity and forensics fields, after DNA identification analysis. The 16 STR loci D3S1358, TH01, D21S11, D18S51, D5S818, D13S317, D7S820, D16S539, CSF1PO, vWA, D8S1179, D2S1338, D19S433, TPOX & FGA, and the locus amelogenin can be amplified simultaneously using the the Identifiler kit (Applied Biosystems, Foster City, CA, USA). This paper presents allele distribution data in the Mexican population of the State of Chihuahua, Northern-Central part of the country. Whole blood was obtained from 161 unrelated volunteers and spotted on FTA paper (Whatman, USA); all individuals were actual residents along the state of Chihuahua, Mexico. Extracted DNA samples were amplified at the 16 loci using the Identifiler kit. Samples were analyzed using the ABI Prism™ 310 Genetic Analyzer (PE Biosystems, Foster City, CA) according to the manufacturer’s recommended protocol. All 15 loci are highly polymorphic in the Mexican sample population with the locus TPOX (55.3%) having the lowest observed heterozygosity, and the locus D2S1338 (88.8%) displaying the highest heterozygosity. The most discriminating loci were D18S51 (PD=0.969) and FGA (PD=0.968). The combined probability of exclusion for the 15 STR loci is 0.99999926. There was little evidence for departures from HardyWeinberg expectations (HWE) in this sample population. Based on the exact test, the loci that departed significantly from HWE were D31358 and D13S317. After employing the Bonferroni correction for the number of loci analyzed, these observations are not likely to be significant. An inter-class correlation test analysis was performed to detect any correlations between alleles at any of the pair-wise comparisons of the 15 loci. A resume of the PD and PE are shown in the following table: 81 Locus PD (Obs) PD (Exp) PE 1 D8S1179 2 D21S11 3 D7S820 4 CSF1PO 5 D3S1358 6 THO1 7 D13S317 8 D16S539 9 D2S1338 10 D19S433 11 vWA 12 TPOX 13 D18S51 14 D5S818 15 FGA 0.92805062 0.94649126 0.91709425 0.85112457 0.91007291 0.90675514 0.93368311 0.92789630 0.95675321 0.92889935 0.90289726 0.79086455 0.96909841 0.85405656 0.96801821 0.92517508 0.95136413 0.91988693 0.86167114 0.91709457 0.91223821 0.94236962 0.93190281 0.96159279 0.94148490 0.91729619 0.79308289 0.97232966 0.88179848 0.97219246 0.59304848 0.66976540 0.57759331 0.45289261 0.57058241 0.55668406 0.64004831 0.60855433 0.70663845 0.63921864 0.57252133 0.36674141 0.75147290 0.49866824 0.75055161 Total >0.99999999 >0.99999999 0.99999926 In conclusion, a Mexican – Chihuahua database has been established for the loci D3S1358, TH01, D21S11, D18S51, D5S818, D13S317, D7S820, D16S539, CSF1PO, vWA, D8S1179, D2S1338, D19S433, TPOX & FGA. All loci are highly polymorphic. The application of the product rule is valid for estimating the rarity of a multiple loci profile for these 15 loci. with these cleaners. Some of these substrates also were treated with either latex or oil based paint in order to determine the ability of the process to detect blood covered by paint. Some substrates also were exposed to environmental conditions for a period of seven days in order to determine potential effects on KPS fluorescence. All of the substrates also were tested using the presumptive tests that employ phenolphthalin or Hemastix strips in comparison with KPS Fluorescein. It was determined that KPS Fluorescein and phenolphthalin sensitivity are comparable, but KPS Fluorescein exhibited cross-reactivity with some cleaning products. Hemastixs were observed to have the greatest sensitivity for blood detection, but exhibited a high frequency of cross-reactivity with a variety of substances. KPS Fluorescein consistently detected blood under latex paint on wood. These substrates were retested at later intervals (i.e., 3 months and 6 months) after KPS Fluorescein treatment, and continued fluorescence was observed at these later times. Both cuttings and swabbings were collected from these samples and processed by DNA extraction and subsequent STR analysis. It was determined that samples that were cut, rather than swabbed for DNA extraction, yielded higher levels of DNA and complete STR profiles. It was determined that KPS Fluorescein does not alter the ability to obtain DNA profiling results. Fluorescein, Blood, DNA STR, Identifiler®, Mexico B103 Obtaining Typable DNA From Bloodstains That Serologically Test Negative B102 A Comparison of KPS Fluorescein to Other Presumptive Blood Identification Techniques and Its Effects on PCR Based DNA Analysis Methods Katie L. Coy, BS*, and Kristen E. Lewis, BS, Virginia Commonwealth University, 1000 West Cary Street, PO Box 842012, Richmond, VA 23284; Ashlee Fulmer, MFS, and Amy Hudson, MFS, The Bode Technology Group, Inc., 7364 Steel Mill Drive, Springfield, VA 22150; and Tracey Dawson Cruz, PhD, Virginia Commonwealth University, 1000 West Cary Street, PO Box 842012, Richmond, VA 23284 Heather J. Seubert, BS*, and Rhonda Craig, MS*, FBI, DNA Analysis Unit I, Room 3230, 2501 Investigation Parkway, Quantico, VA 22135; Jeffrey Leggitt, MS, FBI, Chemical-Biological Science Unit, 2501 Investigation Parkway, Quantico, VA 22135; Gabor Patonay, PhD*, Georgia State University, Department of Chemistry, University Plaza, Atlanta, GA 30303-3083; Kyle A. Hoskins, BS*, and Tani G. Watkins, BA*, Michigan State Police, Bridgeport Laboratory, PO Box 608, Bridgeport, MI 48722; Kevin W.P. Miller, PhD, and Rachael Bartholomew, PhD, FBI, Counter Terrorism and Forensic Science Research Unit, FBI Academy, Quantico, VA 22135; and Richard Guerrieri, MS, FBI, DNA Analysis Unit I, Room 3230, Quantico, VA 22135 The goal of this presentation is to make the forensic community aware that typable DNA may be extracted from bloodstained material even when negative presumptive and/or confirmatory test results are obtained. This presentation will impact the forensic community and/or humanity by changing collection and screening protocols and/or interpretation guidelines for biological evidence testing to assure that valuable forensic evidence is not overlooked. The goal of this presentation is to make the forensic community aware that typable DNA may be extracted from bloodstained material even when negative presumptive and/or confirmatory test results are obtained. This research project was undertaken to determine if typable loci could be obtained from washed and treated bloodstains and how these results compare to serological results, specifically with the ABAcard HemaTrace confirmatory test. Human blood was diluted down to a concentration of 1:500 (undiluted, 1:20, 1:100, 1:250, 1:500), applied to a cotton t-shirt, and then subjected to treatments of machine washing in water, Tide, or bleach. Each sample was then tested using the confirmatory ABAcard test. DNA was extracted by traditional organic methods (phenol/chloroform) followed by quantification of human DNA by the Quantiblot method. Multiplex STR amplification was performed using the ABI AmpFlSTR® Profiler Plus PCR kit. The resulting PCR products were separated and detected by capillary electrophoresis on the ABI 3100Avant Genetic Analyzer and data analysis performed using ABI GeneScan and Genotyper software. The untreated whole bloodstains and undiluted Tide-treated bloodstains were the only samples to produce a positive reading for hemoglobin with the ABAcard test. All other treated and untreated diluted bloodstains gave a negative ABAcard result after washing. However, previously published reports suggest that the ABAcard tests are quite sensitive and possibly able to test positive for whole blood or bloodstains that have been diluted down to 1:1,000,000. Thus, it is noted that traditional Attendees will gain an awareness of another chemical enhancement tool for detecting blood stains and its effects on PCR Based DNA analysis methods. This presentation will impact the forensic community and/or humanity by providing another tool to the forensic community when searching for blood stains. This study was designed to examine the following factors involved with processing evidence for blood staining: 1) to determine if KPS Fluorescein is more sensitive for blood detection than other presumptive tests, 2) to determine if KPS Fluorescein will result in false positives by cross-reacting with various substrates or cleaning products, and 3) to determine if the use of KPS Fluorescein will adversely affect the quality of STR-based DNA profiles developed subsequently. Dilutions of blood were applied to substrates such as dry wall samples, treated and untreated wood samples, and carpet samples. Some of these substrates were then cleaned with typical household cleaners such as Joy Dishwashing Detergent and Clorox Bleach to determine the ability of the process to detect blood as well as assess potential cross reactivity 82 * Presenting Author washing procedures (regardless of treatment) can significantly alter the hemoglobin molecule such that negative results may be shown even when stain extracts actually contain human blood. Results from the DNA profiles indicated that typable DNA was present in many of these treated and diluted bloodstain samples that initially tested negative using the ABAcard confirmatory test. While negative ABAcard results were obtained for all but two stains (undiluted, untreated and undiluted Tide-treated bloodstains), complete or partial STR profiles were obtained from all untreated stains (all dilutions) and several of the treated stains (bleach or Tide). In washed, untreated whole bloodstains, complete profiles were easily generated. In addition, partial profiles (>50% of STR loci successful) were obtained from untreated bloodstain samples that had been diluted down to 1:500. Interestingly, complete and/or partial profiles were also generated from the Tide-treated samples that were either undiluted or diluted down to 1:20. Furthermore, the undiluted bloodstains that were treated with bleach yielded a near complete profile, generating successful results at an average of 89% of all STR loci tested despite the fact that the confirmatory blood serology test on those bleach-treated samples had yielded negative results. This was a significant and unexpected finding because bleach is thought to severely denature most biological molecules, including DNA. Many additional trends were noted with the data generated from these experiments. As one may anticipate, the severity of DNA degradation increases with harsher treatment and along with that, the heterozygote balance decreases, the peak intensity diminishes, and the larger alleles are more prone to allelic dropout. The data produced from these experiments show that confirmatory serology tests are not always reliable predictors of successful STR amplification. Because confirmatory tests such as the ABAcard may often be used in the field and the laboratory to determine what biological evidence to collect and submit for DNA analysis, a negative reading could (in some labs) prevent DNA analysis and thus result in lost forensic evidence. This research demonstrates the value of performing multiple serological screening tests prior to DNA analysis, as well as cautiously interpreting negative results, as typable DNA may be present at trace levels even when the serological protein marker is present at levels below the limits of detection for that test. Further research could investigate amplification of these treated samples using methods designed to detect low copy number DNA since these samples have limited quantities of DNA that vary in quality, similar to what is commonly seen in forensic case samples. DNA, STR, ABAcard HemaTrace B104 Decontamination of Bacterial Spores Without Affecting DNA Molecular Analysis James M. Robertson, PhD*, Douglas L. Anders, PhD, and Douglas J Beecher, PhD, FBI Academy, Laboratory Building, Quantico, VA 22135; and Julie Nicholson, MFS, Donia P Slack, MS, Francene Basalyga, MS, and Robert A Bever, PhD, The Bode Technology Group, 7364 Steel Mill Drive, Springfield, VA 22150 After attending this presentation, attendees will realize that protocols can be used to kill bacteria without destroying the ability to molecularly analyze the DNA contained within the organism. This presentation will impact the forensic community and/or humanity by giving attendees a validated procedure for decontamination without destroying the DNA. The value of this approach is to preserve evidence while allowing clean up in cases where bacteria may be present. In addition, if decontamination has taken place in a rogue laboratory, there may still be evidence that can be evaluated for its DNA signatures. The evidence should not be discarded in such cases. Following the accidental or intentional release of pathogens, surfaces become contaminated and must be treated with a chemical agent to kill the organisms. In rogue laboratories, where pathogens are cultured for criminal or terrorist acts, the perpetrators may decontaminate surfaces and * Presenting Author equipment used to grow the organisms in order to destroy evidence. Although the non-viable pathogens cannot be characterized by their growth characteristics following decontamination, the question remains whether the DNA inside the cells can still be utilized in assays for identification and attribution. Thus, there is a requirement to determine the effects of common decontamination procedures on the activity of DNA in downstream analyses. Research was performed to characterize the effect of several decontamination procedures on DNA as determined by PCR and restriction enzyme analysis. Bacterial spores of Bacillus cereus, a simulant for B. anthracis, were chosen for the studies, because these are very challenging organisms to kill and thus require rigorous decontamination procedures to effect a standard kill of 6-logs in concentration. Suspensions of organisms and dried deposits of the organism on various materials and surfaces of forensic relevance were examined. Manufacturer’s suggested protocols, CDC (paraformaldehyde), and EPA (bleach) protocols were followed to kill the organisms. The materials examined included tile, wood, carpet, cubicle cloth, and painted dry wall. Real-time PCR and pulsed field gel electrophoresis (PFGE) were performed to evaluate the effects on both amplification and restriction digestion, respectively. It was found that 10% bleach was effective in killing the spores at both pH 7 and pH 11, but the matrix on which the organisms were deposited significantly affected the ability to kill the spores. Actril (a mixture of peroxyacetic acid, hydrogen peroxide, and acetic acid), a commercial agent used in hospitals, was as efficient at killing spores as bleach. Actril worked well on all the surfaces and materials tested. DNA extracted from spores exposed to Actril can be amplified more efficiently and consistently than DNA extracted from spores exposed to bleach. PFGE did not reveal differences between the untreated control and Actril-treated spores. Paraformaldehyde (which is converted to formalin gas during the procedure) was very effective, but the DNA was minimally useful for molecular analyses. L-gel, a decontaminating agent reported to be capable of destroying both chemical and biological warfare agents, was not very effective on the spores when the agent was spread over the sample. DNA could be subsequently amplified from the recovered organisms treated with L-gel, but it could not be ruled out that this DNA was derived from cells that remained viable. Sterilox could kill the spores, but the efficacy of this agent was highly affected by the matrix on which the spores were deposited. The DNA was not damaged by Sterilox treatment of the spores. Actril and bleach are the most effective sporicidal agents, and Actril has the least effect on the molecular assays. A validated protocol for the detection of bacteria collected from different surfaces that have been decontaminated by a variety of processes will be presented. Decontamination, Nucleic Acid Amplification/Digestion, Pathogenic Microorganisms B105 Enzymatic-Mediated Digestion of Cellulose for Enhanced Cell Elution for DNA Analysis Jessica C. Voorhees, MSc*, Jerome P. Ferrance, PhD, and James P. Landers, PhD, University of Virginia, Department of Chemistry, McCormick Road, Charlottesville, VA 22904 The focus of this project is the development of an improved method for the elution of cells from a cotton swab taken from a sexual assault victim. The procedure incorporates enzymes for the digestion of the cellulose matrix, resulting in the removal of intact cells, in an effort to circumvent conventional differential extraction. Genetic analysis of perpetrator and victim mixed profile DNA samples obtained from vaginal swabs is a well-established technique in the investigation of sexual assault and rape cases. Unfortunately, the procedures involved in a typical forensic DNA analysis require a great deal, of laboratory time dedicated to a single case, particularly in the sample preparation steps. Because of the time and funding constraints involved in the 83 investigation of such cases, a significant backlog exists in many largevolume DNA analysis laboratories. The current protocol used by law enforcement agencies for recovery of cellular materials from a cotton matrix involves a great deal of sample handling, which directly increases the chances of sample contamination and human error. Furthermore, it is a time-consuming process often requiring overnight incubation of a swab sample for optimal DNA recovery. The extraction solution used in the recovery of DNA from swabs includes proteinase K in the presence of SDS, a combination that selectively lyses the fragile epithelial cells while eluting sperm cells intact. The solution is then centrifuged to pellet the sperm cells, removing them from the solution containing the victim DNA, allowing independent genetic analysis of male and female DNA. The time required for forensic DNA analysis can be greatly reduced by performing the electrophoretic separation on microfabricated glass devices. The speed and efficiency of microchannel separations are due largely to the increased surface to volume ratio of the etched-channels over conventional slab gels. In addition, these devices allow for the integration of additional processing steps, including sample preparation methods. Because centrifugation on a microchip is not trivial, a microchip method for isolating separate male and female DNA fractions has been proposed. This method relies on recovery of intact cells from sample swabs, thus a cell-desorption process that greatly reduces extraction time while leaving cells intact would be advantageous for developing genetic analysis on a micro-total analysis system (µ-TAS). Microscopic examination of a cotton swab on which a semen sample had been applied and allowed to dry suggested that sperm retention on the swab was due to adhesion of sperm cells on the surface of polysaccharide cellulose strands. Preliminary studies have shown that cellulase-based enzymes, which digest cellulose, reduce the time required for sperm and epithelial cells to be released from the swab into solution. In an effort to optimize cellular elution conditions, several different enzymes were evaluated both alone and in combination. Sperm and epithelial cells eluted from a cotton swab were counted using a hemacytometer. Results indicate that elution using enzymes improved the recovery of sperm cells without lysing epithelial cells, and enzymatic sperm cell desorption is greater than that seen with current elution methods. Optimum cellular elution conditions using the enzyme cellulase will be presented. In addition, information regarding the development of a receptacle that interfaces a cotton swab sample with a µ-TAS on a microfabricated glass device will be discussed. Cellulose, Elution, Enzyme B106 Development of Multiplexed Microsatellite Markers in Cannabis sativa Heather L. Erek, BS*, Orchid Cellmark, 1400 Donelson Pike, Suite A-15, Nashville, TN 37217; Tariq Mahmood, PhD, Alberta Research Council, Hwy 16A & 75 Street, Vegreville, Alberta T9C 1T4, Canada; and José Almirall, PhD, Florida International University, Department of Chemistry, University Park, Miami, FL 33199 Attendees will learn that there is a genetic test to provide evidence of cooperation in the production of cannabis. The association of cannabis to sellers, growers and buyers could track networks and show common orgin. This presentation will impact the forensic community and/or humanity by broadening the scope of the DNA analyst and give a genetic test to drug chemists. Cannabis sativa L. (marijuana) is the most frequently used illegal drug in the United States. The marijuana plant can be easily identified through morphological examination and chemical analysis; however there is a need for a method to track distribution networks and to compare plants to determine a common source of origin. A genetic test can enable this association and provide evidence of cooperation in production. Microsatellite markers have distinct advantages over other genetic 84 methods. They have multiple alleles at a single locus, they are reproducible between laboratories, they have a high discrimination power and they can be multiplexed. In this project, a set of Cannabis STR primers previously described by the group and a set of STR primers previously described by Gilmore’s group in Australia were combined and multiplexed into two reactions. The primers include nine trinucleotides, one compound trinucleotide and one imperfect trinucleotide. Both hemp (low THC) plants and marijuana (high THC) plants from different regions of the United States and multiple countries will be tested with the multiplexed primers. The previous studies using these microsatellite markers were able to distinguish clones from non-clones. Efforts are underway to construct a comprehensive genomic map of Cannabis sativa, where the positions of these micorsatellite loci on various chromosomes/linkage groups could be defined. The goal of this project is to determine the level of polymorphism, to show the usefulness for DNA fingerprinting and to measure the genetic relationships between different Cannabis plants, all using a practical set of reactions that can be performed in an operational forensic laboratory. Cannabis Sativa, Multiplexed Microsatellites, STR Primers B107 Evaluation of Track-Etch Filters for Isolating Sperm DNA in Rape Kits Carll Ladd, PhD, Eric J. Carita, MS*, and Elaine M. Pagliaro, JD, Connecticut Forensic Laboratory, 278 Colony Street, Meriden, CT 06451; Alex Garvin, PhD, Bureco Corporation, 18 Gewerbestrass, Allschwil, CH-4123, Switzerland; Timothy M. Palmbach, JD, University of New Haven, 300 Orange Avenue, West Haven, CT 06516; and Henry C. Lee, PhD, Connecticut Forensic Laboratory, 278 Colony Street, Meriden, CT 06451 The goal of this presentation is to evaluate the efficacy of track-etch filters as an alternative approach to the standard selective lysis protocol for isolating sperm DNA profiles from sexual assault samples This presentation will impact the forensic community and/or humanity by providing the forensic community with a tool to evaluate the effectiveness of track-etch filter for the processing of rape kit evidence. The large number of unprocessed sexual assault cases nationwide constitutes an ongoing concern for the forensic community. Many of these cases have sufficient numbers of sperm to generate DNA profiles that could be used to query the CODIS database and identify rape suspects. The standard method for purifying sperm from these swabs is to first resuspend the sample and to selectively digest the epithelial cells with Proteinase K. The intact sperm are then separated from the contaminating solubilized DNA by centrifugation, careful removal of supernatant, and extensive washing of the sperm pellet, all steps that are difficult to automate. The authors have evaluated a vacuum driven filtration method as an alternative approach for separating sperm from digested epithelial cells that is more easily automated in a 96 well format. First, the sample is digested with Proteinase K for 1 hour at 56oC (in standard DNA extraction buffer). Sperm are collected on 2 micron track-etch filters, while the epithelial cell DNA is collected in the filtrate (vacuum pressure = 300 torr). The filters are then washed, and the sperm DNA is solubilized with a reducing agent and collected in the filtrate. The goal of this project is to optimize and validate a faster, more effective, less-labor intensive, and more cost-effective method to isolated sperm DNA from sexual assault samples to address the backlog of unprocessed biological evidence. Mock body fluid sample mixtures (5,000-100,000 sperm per swab) were processed by the vacuum filtration method and the standard differential extraction procedure to determine their ability to separate the male profile from the female profile using the Profiler Plus and COfiler STR kits from Applied Biosystems, Inc. Various parameters were tested to optimize the filtration method including efforts to overcome the recurring problem of filter clogging. These efforts included introducing centrifugation and filter * Presenting Author washing steps, using 10 micron pre-filters to remove cell debris, prewarming washing reagents, using different reducing agents (BME and DTT, various concentrations), adding additional Proteinase K, and slowly adding the sample to the track-etch filter. Membrane clogging can be overcome by centrifuging (3000 G) the sample through the membrane in lieu of vacuum filtration. However, both approaches are less sensitive than the standard differential method and the centrifugation steps are not easy to automate. The authors have shown that use of track-etch filter can be effective for identifying the DNA profile of the semen donor from mixed body fluid samples. To date, the efficiency of separation using track-etch filter has been variable and the method is not as sensitive as the standard differential procedure. This project has been funded by the National Institute of Justice, Grant #2003-IJ-CX-K103 B109 “Taking a Bite Out of Crime”: STR Typing of Biological Evidence Left on Food Remnants Ismail M. Sebetan, MD, PhD*, and Heather A. Zarsky, MFS, Forensic Sciences Program, National University, 11355 North Torrey Pines Road, La Jolla, CA 92037 Attendees will learn of a single STR locus, SE33, that can be used as a DNA-based screening method for heavily bloodstained evidence. A single STR locus such as SE33 can be used to screen heavily bloodstained evidence thus greatly reducing the number of samples requiring a full 16 loci profile. This presentation will impact the forensic community and/or humanity by providing a faster and more cost effective method for forensic laboratories. Forensic laboratories typically identify biological fluids and then, if warranted, the analyst performs DNA analysis to generate a 16 loci genetic marker profile. In most cases this is the most efficient way of processing the case; however, in cases where a large amount of bloodshed has occurred it is time consuming, costly, and it may not be necessary to generate a full 16 loci profile for every stain. In these types of cases it is often necessary to test numerous stains to find the one most probative stain for DNA testing. A faster and more cost effective method would be to screen heavily bloodstained evidence with a single DNA locus to determine if the stain is probative and warrants the full 16 loci testing. HUMACTBP2 (SE33) located on chromosome 5 or 6 is a highly polymorphic STR locus (AAAG repeat sequence) with 32 alleles in the range of 202-323 bp and is one of the loci included in the European STR database, EDNAP. SE33 has numerous microvariants; therefore, it is crucial to fine tune the procedure with regards to the proper gel concentration needed to allow the 1-bp separations. In this study SE33 (Promega’s kit) was extensively validated utilizing the ABI 377 and ABI 310 instrumentation. One hundred buccal swabs were collected and extracted with Chelex, quantitated with QuantiBlot, amplified on an ABI 2700 using 10/20 cycles, and separated on the 377 and 310. This validation included reproducibility/precision, sensitivity, mixture studies, past proficiency samples, and environmental studies. The reproducibility portion of the validation consisted of within-gel and between-gel analysis. The sensitivity study included several sets of samples in the range of 0.03125ng to 5ng. The SE33 types obtained from samples in five proficiency tests were compared to results obtained on the FMBIO II. The mixture study included several mixtures with the following ratios: 1:0, 1:1, 1:2, 1:4, 1:9, and 1:19. In the environmental study ten different blood samples were applied to five substrates (denim, leather, silk, cotton, polyester blend) and exposed to a full year, including rain and snow, of northeastern Pennsylvania weather. The results presented will include the finetuned procedure and a comparison of samples analyzed on the 377 and 310. Future work in this validation will include analysis of samples from nonprobative/no analysis casework. The SE33 types obtained from the samples used in this validation will also be verified at a different laboratory. The goal of this presentation is to demonstrate the amenability of bitten foodstuffs to DNA profiling using current STR methodology and identifies some of the limitations and problems that may be encountered when dealing with food evidence. This research will provide an evaluation of the STR profiles obtained from different types of food possibly encountered at crime scenes and a guide for crime scene investigators and police officers on what kind of evidence should not be overlooked at a crime scene. This presentation will impact the forensic community and/or humanity by showing that DNA typing from bitten foodstuffs is an efficient and reliable alternative source for biological evidentiary samples, and identifies some of the limitations and problems that may be encountered. These results can be used to educate crime scene investigators and police officers on what kind of valuable evidence should not be overlooked at a crime scene. The advancement of DNA technology has increased the capability of DNA profiling to a level in which its greatest boundary is innovation. It is well known that the common biological fluids encountered at crime scenes provide ample DNA for testing. However, in many cases analysis becomes challenging due to the direct deposit of DNA on an unusual surface or item. Bite marks are a piece of evidence that offer an odd, yet fairly common mode of DNA transfer when dealing with burglary, larceny, robbery, and at times, more serious and violent crimes. The number of cases involving bitten foodstuffs is increasing, suggesting a need to look into the DNA profiling of this type of evidence. A potluck was staged as a mock crime scene in which foods such as pizza, corn on the cob, chicken wings and ribs, bite size Hershey’s peanut butter cups, cheese, apple slices, and carrots were partially consumed and remnants discarded. The unconsumed food commodities were swabbed for biological evidence, analyzed for cellular material by microscopy followed by organic extraction, concentration and quantification by slot blot. The AmpFlSTR® Profiler Plus™ PCR Amplification kit was used for all PCR reactions. Samples were concentrated to a final volume of 25 l of which 20 l was used in the amplification reaction. Amplicons were electrophoretically separated using the ABI Prism 310 Genetic Analyzer and analysis of raw data and calling alleles carried out by GeneScan® and Genotyper® softwares respectively. The research results showed that high quality DNA could be extracted with successful STRs typing was obtained. Analyses of the profiles outcomes will be demonstrated. The presence of nucleated epithelial cells in preliminary microscopic results suggested that typing results could be obtained from bitten food products. It was indeed seen that full DNA profiles (9 STR loci + Amelogenin) were obtained from 43% of swabs collected, between 69 loci was seen in 33% of samples, 16% showed 1-5 loci and only 8% failed to type at all. Profile variation based on food-type dependent factors (portion size and eating style, food preparation and natural composition, and swabbing location), are also discussed. Poor profile quality manifesting as allelic dropout is attributed predominantly to PCR inhibition. Degradation and stochastic effects are also considered as sources of allelic dropout but only minimally. Mixed profiles as a ramification of improperly packaged samples are also addressed. In conclusion, this research has largely shown that STR typing of food can be carried out in a highly successful manner, therefore when food evidence is available, DNA results should definitely be sought. Limitations due to low DNA yield and presence of PCR inhibitors can be overcome to produce high quality DNA profiles. Since cross transfer of cellular material was readily evident in a number of samples, any and all occurrences of food items touching must be noted while applying proper packaging protocol. SE33, HUMACTBP2, DNA Bite Mark, STR Typing, Foodstuffs Track-Etch Filters, Sperm DNA Isolation, Differential Extraction B108 Validation of SE33: A DNA-Based Screening Method Carol J. Ritter, MS*, Lawrence Quarino, PhD, Kristen Johnson, BS*, Jennifer Sears*, Robin Shick*, and Katherine Henkelman*, Cedar Crest College, 100 College Drive, Allentown, PA 18104 * Presenting Author 85 B110 Quality and Throughput From a Highly Integrated, Automated Casework Sample Processing Platform Benoît Leclair, PhD*, Timothy D. Kupferschmid, MFS, Corey L. Schwensen, MS, Stephen Gresko, BS, Victor Thompson, BS, Brian E. Ward, PhD, and Tom Scholl, PhD, Myriad Genetic Laboratories, Inc., 320 Wakara Way, Salt Lake City, UT 84108 Te attendee should have a clear understanding of what current automated sample processing technologies can do for current casework loads and backlogs, in terms of process quality, flexibility and throughput. Currently, long manual processing times give offenders ample opportunity to re-offend. Highly integrated automated solutions have the potential to eliminate backlogs, provide quick turn-around on newly submitted cases, increase crime resolution rates, and deny repeat offenders the opportunity to re-offend. Countless victims of violent crimes could be spared their ordeal. The demanding nature of forensic casework evidence makes of sample processing a labor-intensive manual undertaking. Submissions of criminal casework involving biological evidence currently exceed installed processing capacity in numerous jurisdictions. It is currently estimated that across the United States, over 500,000 criminal cases with biological evidence are awaiting processing. Highly integrated, automated biological specimen processing platforms have been in use in high-throughput clinical and large-scale genetic research facilities for many years, and more recently in felon genotype data banking initiatives. These systems have reached a level of maturity that warrants leveraging this experience into the casework sampleprocessing arena, despite the demanding nature of casework biological evidence. This report presents the result of the development efforts towards the implementation of highly integrated, automated casework sampleprocessing platform. The development goals were to achieve performance standards comparable to or better than a traditional manual process with regards to genotyping results, sample consumption, process flexibility, and chain-ofcustody. In addition, the platform fully exploits process tracking, throughput and scalability capabilities. The entire process is overseen by a custom-built LIMS addressing the specific requirements of forensic casework. It tracks samples, their derivatives and associated process-generated data from the moment the samples enter the facility. It controls all robotic processes, ensures process integrity and sample tracking through the extensive use of barcodes. It automatically computes quality metrics and performs trend analysis to monitor / document process quality. Experienced analysts manually search exhibits for biological stains. According to the specifics of the case, cuttings enter the automated processing platform under the “Differential” or “Non-Differential” pathways. TECAN robotic liquid handlers perform all processes from cell lysis to the production of capillary loading plates. Robot-based chemistries are optimized to deliver quantitative DNA extraction yields from scarce samples. DNA is extracted with ChargeSwitch para-magnetic beads, and quantification is performed with the Quantifiler Autosomal and Y kits. All commercial STR megaplexes are supported. The high precision, accuracy and reproducibility of robotic pipetting contribute significantly to overall process quality. The batching of samples for robotic processing yields considerable efficiencies in process execution time. To support process flexibility and minimal exhibit/extracted DNA consumption, the LIMS provides the analyst with customizable re-work pathways from any given step within the platform, as well as process branch-off points to return a sample to offline processing. The LIMS incorporates numerous user interfaces that allow the analyst to follow / audit the processing path of a sample and maintain total control on the processing of samples. The entire system is built under a novel architecture that allows for new processes or submitting agency preferences to be quickly integrated, extensively tested and validated. 86 A complete developmental validation study was conducted according to SWGDAM guidelines and demonstrated the reliability of the platform. This presentation will showcase case examples processed by this high quality, automated system. Automation, Casework, Validation B111 A Concordance Study Comparing Different Amplification Chemistries and Electrophoretic Platforms for a Databasing Program G. Sue Rogers, MSFS*, Alabama Department of Forensic Sciences, 501 12th Street South, Birmingham, AL 35233; Bridget Tincher, MSFS, Marshall University, Forensic Science Center, 1401 Forensic Science Drive, Huntington, WV 25701-3628; Angelo Della Manna, MSFS, Alabama Department of Forensic Science, 501 12th Street South, Birmingham, AL 35233; Terry Fenger, PhD, Marshall University, Forensic Science Center, 1401 Forensic Science Drive, Huntington, WV 25701-3628; and Rhonda K. Roby, MPH, Identity Quest, LLC, 13203 Astoria Hill Court, Suite J, Germantown, MD 20874 The goal of this presentation is to present the results of an extensive concordance study and collaboration performed by the Alabama Department of Forensic Sciences (ADFS) and Marshall University Forensic Science Center (MUFSC) in order to address the State of Alabama’s backlog of convicted offender samples. This presentation will impact the forensic community and/or humanity by demonstrating that either multiplex reagent kit or instrument platform can be effectively used to process convicted offender samples. The study demonstrates that the procedures employed are robust and valid. Furthermore, this study will present recommendations for the forensic community regarding the interpretation of DNA profiles developed from convicted offender samples and their subsequent entry into CODIS in order to maximize the current searching algorithm to aid in the identification of the perpetrators of violent crimes. Concordant results using different fluorescent-based STR (short tandem repeats) genotyping systems for the analysis and databasing of convicted offender samples are critical for accurate matching to nonsuspect casework and to link serial crimes. In order for a state to upload its database of convicted offender DNA profiles into national level of the COmbined DNA Indexing System (CODIS), each profile must be comprised of the standard 13 core STR loci. Prior to this requirement, the ADFS had generated a convicted offender DNA database with eight (8) STR loci of the 13 core STR loci. With the support of the National Institute of Justice, the Alabama Department of Forensic Sciences (ADFS) and Marshall University Forensic Science Center (MUFSC) initiated a collaborative study to conduct additional DNA testing on these convicted offender samples to develop results for the remaining five (5) core STR loci and to demonstrate concordance of the STR results obtained, regardless of the testing platform employed. The STR results submitted by MUFSC were generated using System 1, the ABI PRISM® 3100 Genetic Analyzer, the AmpFISTR® Identifiler® PCR Amplification Kit, ABI PRISM® GeneScan® Software version 3.7 and Genotyper® Software version 3.7.1 (Applied Biosystems, Foster City, CA). The STR results from samples previously processed by the Alabama Department of Forensic Sciences were obtained using System 2, the Hitachi FMBIO® II Fluorescent Imaging Device (Hitachi/Miraibio Genetic Systems, Alameda, CA), the PowerPlex® 1.1 System (Promega Corporation, Madison, WI), and FMBIO® Analysis and STaR Call ™ (Hitachi Genetic Systems). This study demonstrates that the combination of instruments, reagents, procedures, and analyses employed by both of these institutions provided concordant and accurate genotyping results for databasing purposes. * Presenting Author Beginning in April 2002, the MUFSC submitted 5,000 STR profiles to the ADFS using System 1 had previously been processed using System 2 by the Alabama Department of Forensic Sciences. Overall, there is incredible concordance for all 5,000 samples between the two (2) systems. Greater than 99.98% concordance is achieved when comparing the overlapping eight (8) loci present in both systems. Any differences noted have been identified and summarized into the following categories: lower resolution when using System 2; stochastic amplification; and allelic dropout. An example and respective percentages of each category will be presented. Resolution with the capillary electrophoresis platform is much better than resolution using slab gel electrophoresis. Further, it is known that the amplification kits from various manufacturers incorporate dissimilar different primer sets which may result in differences in the amplification conditions and may lead to destabilization of the amplification and eventually stochastic effects and/or allele dropout. The frequency of stochastic effects and allele dropout is extremely low. The manufacturer of the kit with the loci exhibiting the highest frequency of allelic dropout observed in this study (i.e., D13S17 and D16S539) has subsequently modified PowerPlex® kit(s) to incorporate amplification of the null alleles. Rare observations of stochastic effects and/or allelic dropout were observed and documented at various loci in both testing systems. The searching algorithms available within the CODIS software which employ moderate stringency searches and allow for a selected number of mismatches can address any low frequency differences between testing platforms. This concordance study demonstrates that either multiplex reagent kit or instrument platform can be effectively used to process convicted offender samples. The study demonstrates that the procedures employed are robust and valid. Furthermore, this study will present recommendations for the forensic community regarding the interpretation of DNA profiles developed from convicted offender samples and their subsequent entry into CODIS in order to maximize the current searching algorithm to aid in the identification of the perpetrators of violent crimes. Supported under Award Number 2001-RC-CX-K002 from the Office of Justice Programs, National Institute of Justice, Department of Justice. Points of view in this document are those of the authors and do not necessarily represent the official position of the U.S. Department of Justice. CODIS, Concordance, Databasing B112 “Safe in the Crosshairs”: One-Way™ Bullet Resistant Glass Susanna Rudy, RN, MSFS*, University of California Medical Center at San Diego, 200 West Arbor Drive, San Diego, CA 92103 After attending this presentation, law enforcement, investigators, ballistics specialists, and criminalists will be informed of this unique type of glass, who produces it, how it is manufactured, how it works, the ammunition that it repels and the types of individuals and organizations that have access to this technology and are currently using this product as well as others offered by Labock Technologies in order to protect against terrorism and criminal activity. An awareness of the existence of this type of glass has a significant impact on the way law enforcement and various other governmental agencies may chose to outfit their personnel for security purposes. This presentation will impact the forensic community and/or humanity by benefitting to criminalists and ballistic specialists in solving forensically significant cases involving such glass. As the World focuses its attention on the sad demonstration and threat of terrorism and increasingly more violent crimes, hopes for safety and security lie in the skills and intelligence of science and technology. They are the minds that work quietly behind the scenes to develop state of the art deterrents and safety devices necessary to protect the lives of those who speak for a greater World peace and stay one step ahead of those who mastermind to destroy it. * Presenting Author Labock Technologies Inc., a research, development and manufacturing company known to many government agencies and corporations for its armoring technologies has come to the forefront in the war on fighting terror by introducing a revolutionary new concept in a bullet proof glass design. One-Way™ bullet resistant glass. Labock is the only company that has successfully produced a bullet resistant, high clarity/glass/plastic armor system that offers protection against hostile fire while allowing successful return fire from within the confines of the vehicle or structure without damaging the integrity of the glass. The glass is made of a series of polycarbonate, acrylic and glass layers merged in a special manufacturing process. It is significantly lighter, has lower distortion and gives up to 90% clarity. Passing all of the standard governmental agencies ballistic tests 100% of the time, the One-Way™ protection system (unidirectional armor) is resistant up to NIJ level III standards. This technology is offered to distinguished clients only. This technology has been featured in several publications including Discover Magazine, New Orleans city Council News and the History Channel. This state of the art technology is being used as the primary source of protection in many vehicles and buildings that are currently in the rebuilding phase of Iraq. Terrorism, One Way (TM) Bullet Resistant Glass, NIJ Level III Standards B113 Effects of Aging on Pressure Sensitive Tape Analysis: A Preliminary Study Rebecca Bucht, BSc*, 309 West 76th Street, #4A, New York, NY 10023 The goal of this presentation is to present preliminary results on how accelerated aging and prolonged exposure to moderate heat affect the IR spectra of the backing and adhesive layer of tapes, as well as microspectrophotometry of the backing. The comparison of tapes to database information and standards often involves the use of IR spectra of the backing and adhesive, as well as MSP of the backing. This presentation will impact the forensic community and/or humanity by showing the important of considering how exposure to different environments can affect these features. Forensic tape analysis involves either attempting to match two pieces of tape or attempting to identify a tape by comparing it to standards in a database. Parameters contained in the database include a number of physical features and chemical properties. Physical features include functions of dimensions and morphology, which are not necessarily stable when the tape is taken off the roll and applied. More stable and objective measurements such as determination of the components of the backing and adhesive, as well as MSP measurement of the color of the backing, have been proposed to add more stable dimensions to the databases. IR spectra of the backing and adhesive have been proposed in order to identify the components of the backing and adhesive layers. However, both the polymers and additives used in pressure sensitive tape backings and adhesives degrade when exposed to a number of factors such as oxygen, humidity, mechanical stress, aggressive media and ionizing radiation and temperature. This degradation has the potential of altering the appearance of the IR and MSP spectra. A selection of tapes was obtained, including office, duct, electrical and packing tapes. The backing layers were analyzed by ATR-FTIR and MSP, and the adhesive layers by DRIFTS. One set of tapes was then exposed to moderately heat in an oven set to 55°C and another to accelerated aging according to the ISO 4892-2 standard. T he tapes were attached to two different plastic films, PVDC and LLD-PE. Spectra of the backing and adhesives were then obtained by ATR-FTRI and DRIFTS and compared to those of the unexposed tape. The main polymer component of the spectra did not change significantly, but noteworthy changes were observed in smaller peaks that could be used to differentiate between tapes of the same base polymer but 87 different manufacturers. As expected, not all types or brands of tapes reacted similarly to the environmental exposure. In addition, some changes in the adhesive spectra appear to be influenced by the substrate the tape was deposited on during its exposure to heat or weathering. Only one set of tapes was exposed to each of the environments, and only one set of analyses performed on each sample, so the results would have to be confirmed by a more comprehensive study. More work needs to be done to validate and further define these results, but it seems clear that effects from exposure to different environments, and potentially substrate effects should be taken into consideration when analyzing pressure sensitive tapes. Pressure Sensitive Tape, IR, Database B114 A Comprehensive Training and Professional Development Program for an Accredited Forensic Laboratory in the Public Sector Gareth P. Jones, MSc, and Kimberley A. Johnston, MSc, BA*, Centre of Forensic Sciences, 25 Grosvenor Street, 2nd Floor, Toronto, Ontario M7A 2G8, Canada The goal of this presentation is to provide a laboratory seeking American Society of Crime Laboratory Directors/Laboratory Accreditation Board (ASCLD/LAB) accreditation with guidance and a framework for implementing a successful Training and Professional Development program. This presentation will impact the forensic community and/or humanity by providing a model that can assist an organization in creating a training and professional development program and policy that meets or exceeds ASCLD/LAB requirements and is considered by all staff as an integral component in their employment. This presentation will describe the design of a comprehensive training and professional development program suitable for an ASCLD/LAB accredited forensic laboratory in the public sector. The relevant ASCLD/LAB principle, standards and criteria are: • 1.3.3 TRAINING AND DEVELOPMENT PRINCIPLE Training and development of employees must be emphasized to improve accuracy, increase productivity, and enable them to assume greater responsibility. • STANDARDS AND CRITERIA A training program to develop the technical skills of employees is essential in each applicable functional area. • 1.3.3.1 Does the laboratory have and use a documented training program in each functional area for employees who are new, untrained or in need of remedial training? (E) • A formalized personnel development program is important to prepare employees to assume more responsible jobs. • 1.3.3.2 Does the laboratory have an employee development program? (I) The Centre of Forensic Sciences (CFS) Training Manager is accountable for ensuring that high-quality, relevant training and education services are made accessible and provided to staff on a regular basis. This work is facilitated through a Training and Education Committee, which consists of a manager from each section/unit (total 11) and is responsible for providing the Training Manager with: • Assistance, advice and support in the development of CFS-wide training initiatives. • Staff and resources for the delivery of CFS-wide training initiatives. • Assistance, advice and support in the development of an annual training plan of external educational commitments. • Staff and resources for the delivery of all external educational commitments. • The development and implementation of internal training program(s) specific to each section/unit. 88 • The training and development plans for each member of their staff. • Assistance in the development and maintenance of a process for the allocation of funds for specific and general training initiatives. • A forum for representatives from each CFS section/unit to discuss general and specific training needs. The Training Unit is responsible for implementation of committee resolutions and decisions. The training and professional development program at the CFS, which meets ASCLD/LAB requirements, is detailed in the lab’s Training and Professional Development policy. Opportunities for training and development are provided to all CFS staff (operational/administrative support staff, technologists, scientists, and managers). The training and development program for all staff commences with the offer of employment, when each new staff member receives an outline of his/her training and development program for the first year. Additionally, each staff member, in conjunction with her/his manager, develops an annual performance and development plan that includes training and development opportunities identified during each individual’s performance review. The manager responsible for training and development in each section/unit compiles the individual learning plans into a section/unit plan and submits it to the CFS Training Manager for organizational integration and implementation within the funding envelope of the Centre’s annual training budget, which is sheltered from erosion. Dispensation of training funds is influenced by many factors including the size of each section/unit, available operational and developmental opportunities, and organizational pressures. The CFS training and professional development budget’s funding is approximately 4 to 6% of the Centre’s salary budget. This level of funding is aligned to the benchmark established by both the Government’s Human Resource Strategy – which outlines that high performance and high technology organizations provide approximately 6% of their salary budget for training – and the American Society of Training and Development’s annual reports of funding levels for leading edge organizations (Training Investment Leaders). New staff complete an orientation program that provides health and safety training, information on the role of the CFS in support of the justice system and public safety, access to Human Resources information and an introductory tour of the facility. Basic training for new staff, in addition to the orientation program, consists of organization-wide and section/unit-specific programs that differ depending on the section/unit and the nature of the individual’s job. Organization-wide training includes some or all of the following elements: Policy training; Quality Assurance, Training, and Library information sessions; Miscarriages of justice training; Court system training; Centre Receiving Office training; and autopsy attendance. Section/unit specific training includes practical and theoretical components and a variety of competency tests. Professional development is based on individual learning plans and often includes consideration of the technical and behavioral competencies identified for the individual’s job, as well as section/unit, organizational, Ministry and Ontario Public Service needs and requirements. Generally, professional development opportunities include: attendance at seminars and courses; attendance at administrative, professional, scientific, or management conferences; membership on inter- or intra-ministry work projects/committees; membership on scientific/technical working groups; cross-assignments or re-assignments; and secondments. In addition, professional development may include behavioral competency assessment in alignment with the Ontario Public Service competency models developed for scientists, technologists and managers at the Centre. Staff may also be provided with opportunities to deliver client education as part of their development program. Training, Professional Development, Accreditation * Presenting Author B115 Potential for Brand Name Identification by Tomato Seed DNA Typing B116 Progress in the Individualization of Gasoline Residues Cheng-Lung Lee, MFS*, Nuclear Science Department, National Tsing Hua University and Hsinchu Municipal Police Bureau, 101, Section 2, Kuang Fu Road, Hsinchu, 300, Taiwan, Republic of China; Heather Miller Coyle, PhD, 278 Colony Street, Meriden, CT 06451; Ian C. Hsu, PhD, Kuang Fu Road, Hsin Chu, 101 Sec 2, Taiwan, Republic of China; and Albert Hooper, PhD, Carol Scherczin, PhD, Timothy M. Palmbach, PhD, Henry C. Lee, PhD, and Albert B. Harper, PhD, JD, Institute for Forensic Science, 300 Orange Avenue, West Haven, CT 06516 J. Graham Rankin, PhD*, Jeremy Wintz, MSFS, and Raymond Everett, BS, Marshall University, Forensic Science Program, 1401 Forensic Science Drive, Huntington, WV 25701 In recent years, DNA has been successfully extracted and analyzed from a variety of plant materials including seed pods, leaves and other vegetative matter from plant species such as Cannabis sativa (marijuana) and other ornamentals (Palo verde, Sutera). The ultimate goal of plant DNA typing is to extend the microscopic analysis of vegetative trace materials to further classify or individualize the sample. Since seeds are small, often adherent to clothing, and edible (stomach contents), they are a good choice to optimize for DNA typing from crime scene evidence. As a model system, the authors are using the amplified fragment length polymorphism (AFLP) method for DNA individualization of Lycopersicum esculentum (tomato) seeds that are common to many cuisines around the world. The results indicate that high quality DNA could be extracted from fresh tomato seeds after they passed through the human digestive system using a commercially available plant DNA extraction kit. Although the sample weight was significantly less than the recommended amount for the kit, high quality and sufficient quantity of DNA was isolated from a single tomato seed to generate an AFLP profile. When DNA profiles were compared from different seeds, several variety-specific markers (DNA fragments) were identified. Although the sample size was small (n = 4 seeds each per 5 different tomato varieties), these markers look promising for tomato variety identification. Additional screening of larger tomato populations is in progress. Not only will new markers be identified and correlated with a tomato variety, but the percentages of shared markers can be estimated by comparisons of samples from the same variety and between varieties. A brief description of tomato varieties and their cultivation history will be provided in this presentation. Tomatoes have been extensively cultivated and many share a common genetic ancestry and so have been notoriously difficult to distinguish genetically due to high levels of inbreeding. The AFLP technology has been specifically developed to generate high marker saturation across plant genomes to overcome the inbreeding issue. In addition, a comparison of fresh and processed tomato seeds and the subsequent recovery of DNA were performed. Fresh tomato seeds, both digested and undigested yielded an average of 62.5 ng/ 50 uL of DNA per seed embryo. Interestingly, DNA was not recoverable from processed tomato seeds from spaghetti sauces or canned tomatoes presumably due to the pressure and heat treatment during processing. Further studies are in progress to define if other forms of cooking (e.g., oven baking, boiling etc.) affect the ability to recover PCR quality DNA from tomato seeds. In summary, the data demonstrate that AFLP analysis is appropriate for individualization of unprocessed tomato seeds and may be useful for linkage of stomach contents back to a location, especially once variety-specific markers have been determined. Forensic Botany, Plant DNA, Tomato Seed After attending this presentation, attendees will have a better understanding of the methodology of gasoline individualization, the problems associated with comparing neat gasoline samples with residues from fire debris and the use of multivariate statistical methods in making those comparisons. This presentation will impact the forensic community and/or humanity by providing better methods for individualizing gasoline residues in suspected arson cases. Gasoline is a frequent accelerant used by arsonists. Identification of gasoline in fire debris is relatively easy even when greatly evaporated. However, when a suspect is apprehended with gasoline residue on his clothes or a gasoline can in his vehicle, the question arises if the gasoline residue from the fire debris can be matched with that found with the suspect. This has proven to be a more difficult challenge. Further recent legal challenges to comparison evidence have stressed the necessity of establishing a statistical probability for that match. Julia Dolan (ATF National Research Lab, Ammendale, MD), at the 2002 meeting of AAFS, presented a high-resolution GCMS method for comparing gasolines based on 20 sequential area ratios of 34 target compounds from 3-methylpentane through the 1-methylnaphthalene. Her data set included 36 different gasolines, including both regular and premium, mostly from around the Washington, DC area. In addition to neat gasolines, 25% and 50% evaporated samples were analyzed. Mark Sandercock (RCMP, Edmonton, AB, Canada) has developed an alternative method for gasoline individualization based on two and three ring polyaromatic hydrocarbons that he was able to differentiate even at 90% evaporation. Neither study involved simulated fire debris. As part of an ongoing study, the authors have assembled a collection of over 100 gasoline samples from across the U.S. and have analyzed many of these by both the Dolan and Sandercock methods. In addition, analyses of simulated fire debris (charred wood and carpet pad) comparing the gasoline residues with their corresponding neat gasoline will be presented. The ASTM method E1412 (activated charcoal strip adsorption, ACS) is typically used in fire debris analysis but discrimination effects have been reported in the past. Effects of the ACS method on comparison of gasolines by both methods will also be presented. Gasoline Individualization, Fire Debris Analysis, Principal Components Analysis B117 Adsorption Saturation and Chromatographic Distortion Effects on Passive Headspace Sampling With Activated Charcoal in Fire Debris Analysis Mary R. Williams, BS, Denise Fernandes, BS, Candice Bridge, BS, Derek Dorrien, Stefanie Elliott, and Michael E. Sigman, PhD*, National Center for Forensic Science, University of Central Florida, P.O. Box 162367, Orlando, FL 32816 Attendees will emerge from the talk with an appreciation for the complicating effects of adsorbent saturation on the resulting chromatographic profile in ignitable liquid analysis. Audience members will be provided with an understanding of the causes of chromatographic distortion and ways to alleviate these effects in arson investigations. * Presenting Author 89 This presentation will impact the forensic community and/or humanity by presenting results which will be discussed along with their relevance to ASTM methods and possible impacts on the analysis of ignitable liquids residues for pattern matching and source identification. The recovery of hydrocarbons from an equimolar test mixture containing C7 – C10 components has been investigated to determine the influence of hydrocarbon vapor phase concentration on the molar quantities and ratios of the components recovered by passive headspace sampling with activated charcoal. In a one-quart container, hydrocarbon volumes as small as 24 µl (liquid) were sufficient to saturate a 99 mm2 activated charcoal square. Hydrocarbon displacement from the saturated surface of the activated charcoal resulted in significant distortion in the molar ratio and chromatographic profile of the recovered hydrocarbons. Hydrocarbon adsorption data for the C7 – C10 equimolar mixture was used to estimate the average surface area of the activated carbon at 1128 ± 197 m2/g. In order to demonstrate the effect of adsorbent saturation on a commonly encountered commercial accelerant, similar experiments were performed with gasoline. Passive headspace sampling of similarly small volumes of unweathered gasoline resulted in significant distortion of the chromatographic profile of the components recovered from the headspace by passive sampling. The chromatographic profile of the recovered hydrocarbons closely resembled 75% weathered gasoline. A method of subsample collection and analysis was demonstrated as a way to alleviate distortion from samples containing heavy loadings of ignitable liquids. Carpet and carpet padding were placed in a quart container and a heavy loading of unweathered gasoline was placed in the center of the carpet and padding. The container was closed and heated to 60ºC for a period of 16 hours to bring the volatile components from the gasoline into the vapor phase. The container was then cooled to room temperature and a subsample of the carpet padding was removed and placed into an empty one-quart container along with a 99 mm2 activated charcoal strip. The subsample was heated to 60ºC for a period of 16 hours and the activated carbon was subsequently extracted with CS2 and analyzed by GC/MS. The resulting chromatogram closely resembled the chromatographic profile of the unweathered gasoline. These results will be discussed along with their relevance to ASTM methods and possible impacts on the analysis of ignitable liquids residues for pattern matching and source identification. Fire Depris, Ignitable Liquids, Trace Evidence B118 Application of a Standardized Gas Chromatography Tandem Mass Spectrometry Method for the Improved Detection of Ignitable Liquid Residues Jeannette M. Perr, BS*, Kenneth G. Furton, PhD, Marilyn Prieto, and José R. Almirall, PhD, Florida International University, International Forensic Research Institute, Department of Chemistry and Biochemistry, 11200 Southwest 8th Street, CP 316, Miami, FL 33199 The goal of this presentation is to decribe an approach to improve the detection and identification of ignitable liquid residues (ILR) through development of a standardized gas chromatography ion trap tandem mass spectrometry (GC/IT/MS/MS) method that is simple to implement and interpret. This presentation will impact the forensic community and/or humanity by describing an approach to improve the detection and identification of ignitable liquid residues through development of a standard GC/IT/MS/MS method that is simple to implement and interpret. Data in support of the method will be provided and the advantages of the method will be shown through examples where GC/MS analysis alone does not provide conclusive results. This method allows for improved analysis of ILR evidence by overcoming previous disadvantages of MS/MS methods without increased examiner labor or time or the need for complicated interpretation software. 90 The detection and identification of small quantities of ignitable liquid residues in fire debris evidence can be difficult due to: significant alteration of these mixtures during the fire, recovery methods that afford poor selectivity and/or sensitivity, and co-extraction of interfering products (IP). The objective of this project was the development of a standardized GC/IT/MS/MS method that can be employed on an ion trap mass spectrometer, independent of manufacturer. Flexible chromatographic parameters and uniform ionization parameters were incorporated in the creation of a standardized GC/MS/MS method for ILR analysis. Regions of uniform ionization, termed bins, were created using the standard n-alkanes, C8 through C23, as markers. The general approach described in this presentation can be used in any GC/IT/MS/MS instrument and the spectra generated can be shared as a guide between instruments. An examiner can generate his/her library for use in their laboratory or use a previously created IT/MS/MS library due to the uniform ionization. Changes in chromatographic parameters would require minor adjustments of the bins. A searchable mass spectral database produced from this method would facilitate identification of components found in ignitable liquids (ILs) and interfering products (IPs). Chromatograms obtained using the standard IT/MS/MS method will be presented and compared to data generated by typical MS methods for different ILs and their corresponding weathered fractions in the presence of interfering products. Data in support of the advantages of the method will be shown through examples where GC/MS analysis alone does not provide conclusive results. These experiments will improve current analysis of fire debris without significantly changing the workload of the examiner or the time required for analysis. It is anticipated that the results using GC/IT/MS/MS will not require sophisticated training or data analysis programs. This method is expected to allow for the identification of ILRs that might currently go undetected. Ignitable Liquid Residues, Interfering Products, GC/IT/MS/MS B119 Fire Debris Analysis Using GC/MS/MS: Detailed Case Study Interpretations Dale A. Sutherland, BSc, CChem*, Activation Laboratories, Ltd., 1336 Sandhill Drive, Ancaster, Ontario L9G 4V5, Canada Attendees will gain a better appreciation and measure of the increase in specificity gained through the use of the GC/MS/MS method, how it is used, and the benefits of potentially using this method in their practice. The conferee will also gain an insight into recent research published on the use of GC/MS/MS on fire debris. This presentation will impact the forensic community and/or humanity by providing forensic scientists with a mean to better evaluate and consider the importance of the use of GC/MS/MS for the analysis of ignitable liquids in fire debris. These scientists are being asked to vote on the publication of this method as an ASTM guideline and it will afford them the chance to interact and see detailed results from the use of this method as explained by one of the most experienced forensic scientist in the use of this technique. Scientists in this field will then further understand it’s capabilities as a more specific analysis with an associated higher level of confidence in the result and how it applies to presentation in the courtroom. Through ASTM publication this method will reduce the occurance of false negative results and allow this forensic field to keep abreast with the current level of science. The use of GC/MS/MS in the analysis of fire debris is being proposed by an ASTM sub-committee of the Criminalistics section. A guideline has been written, reviewed, revised and is now pending appropriate voting by members. This instrumentation and use of GC/MS/MS is well proven and has been used in other fields of testing such as Environmental, Pharmaceutical and the Drug Testing field for several years. The analysis of fire debris using GC/MS/MS itself has now been published in journals * Presenting Author for over seven years. The use of a second coupled mass spectrometer based segment significantly improves the selectivity of the analysis and is the instrument of choice when presented with the objectives to detect and identify trace contaminants within highly complex matrices. The analysis for ignitable liquids in fire debris fits this situation and GC/MS/MS is the instrument of ultimate choice for such mixture analysis. The advent of GC/MS in this field, through today’s wide use of ASTM E-1618 represented a significant step forward in the reliability of the analysis of fire debris. However, when these samples are highly weathered or contain possible trace quantities of an ignitable liquid in a particularly complex debris sample, GC/MS may not yield confident court presentable results or may not illustrate to the scientist enough ignitable liquid characteristics to determine a confident positive identification. In such circumstances the scientist may often suspect the presence of an ignitable liquid but without sufficient presentable characteristics the result may be undetermined. The use of GC/MS/MS may often more clearly illustrate the required characteristics for the identification of an ignitable liquid when present and thus potentially avoid the reporting of a false negative determination. This method can provide profiles of superior comparability to reference ignitable liquid analyses than with GC/MS as the effects of pyrolysates on the profiles of various compound class characteristics of ignitable liquids can be vastly reduced. The aspect of presentability is becoming more important for the courtroom so that the judge and members of the jury can clearly conclude similarity of the results of fire debris analysis to reference ignitable liquids and thus have more confidence in the result and better information to make their conclusions. The use of GC/MS/MS has already been presented in court specifically for the analysis of fire debris samples. This presentation will focus on the step-by-step interpretation, using a class pattern recognition approach, of two samples from different case studies that will illustrate the specific advantages and limitations of the analysis of fire debris using GC/MS/MS versus the GC/MS method outlined in ASTM E-1618. The presentation will also address previous concerns of sensitivity with reference to the abilities of the trained K9 often used to locate samples at the scene. Analysis, GC/MS/MS, Debris B120 Transfer and Adherence of Gasoline Vapors Onto Clothing in an Enclosed Room Raymond J. Kuk, MS*, Bureau of Alcohol, Tobacco, Firearms, and Explosives, Forensic Science Laboratory, Washington, 6000 Ammendale Road, Ammendale, MD 20705; and Edward C. Diamond, BS, Bureau of Alcohol, Tobacco, Firearms, and Explosives, Ann Arbor, MI Field Office, Ann Arbor, MI 48104 The goal of this presentation is to provide information regarding the exposure of clothing to gasoline vapors in an enclosed room and the subsequent laboratory testing of the clothing. Chemists who analyze fire debris are sometimes requested to examine a suspect’s clothing for the presence of ignitable liquids. The presence or absence of ignitable liquids on a suspect’s clothing may play a significant role in a trial. Several factors may have an effect on the retention of ignitable liquids and or vapors on the clothing. These factors include whether or not liquid was spilled on the clothing, duration of exposure and the time between exposure and collection of the clothing. This study was conducted as part of the ATF certification program for certified fire investigators and examined the transfer and adherence of gasoline vapors to various clothing materials and the effect of time between exposure and collection. The clothing items used in this study were swatches from a pair of 100% cotton jeans, a 100% cotton t-shirt, a 100% polyester t-shirt, and a 50:50 cotton:polyester sweatshirt. In order to simulate a standard room, a 1:12 scale model of a 10-foot by 12-foot room was constructed using plywood, drywall and pressboard flooring. No doors or windows were cut into the walls in order to minimize the escape of vapors. * Presenting Author The interior of the room was not finished in order to minimize the available material on to which the gasoline vapors could adhere. These factors were selected to simulate a worst-case scenario for an enclosed room. Four rooms were constructed - one for each type of fabric. A metal baking pan was placed on the floor of each model to contain the liquid gasoline. Fishing wire and binder clips were used to suspend five three-inch square swatches of each fabric material approximately three inches above the metal pan. It was assumed that one-half gallon of gasoline would be a sufficient amount to pour around a 10-foot by 12-foot room. This volume of gasoline was reduced by a factor of 12 and poured into the metal pan. A drywall ceiling was put into place and the room was enclosed for a period of 15 minutes. After 15 minutes, the ceiling was displaced and the fabric swatches were removed. One swatch was immediately placed into a quart can, and the can was sealed. The remaining swatches were exposed to atmospheric conditions for 15 minutes, 30 minutes, 60 minutes, and 24 hours. After each time frame, one fabric swatch was placed in a quart can and the can was sealed. This process was repeated twice more with fresh gasoline for each trial. A control sample of each fabric was also collected. Each fabric sample was extracted using passive headspace concentration with a charcoal strip (ASTM E-1412). Each sample was heated at 65°C for 16 hours and extracted with 150 µL of carbon disulfide. The samples were then analyzed using a gas chromatograph – mass spectrometer. The total ion chromatogram and the aromatic (m/z = 91 + 105 + 119 + 133) extracted ion profile were used to evaluate the data. All of the control samples were negative for the presence of gasoline. Each fabric sample packaged immediately after exposure showed indications of gasoline in both the total ion chromatogram and the extracted aromatic profile, but the patterns were not significant enough to warrant a positive identification of gasoline. After 15 minutes of atmospheric exposure, only the polyester t-shirt and the sweatshirt showed indications of gasoline in the aromatic extracted profile. After 30 minutes of atmospheric exposure, none of the samples showed any indications of gasoline. This test was repeated with 20 µL gasoline placed on each of the fabric swatches to simulate approximately 12 drops of gasoline spilled on each fabric sample. Positive identification of gasoline (evaporated) on each of the fabric materials could be made on the samples immediately packaged and after 15 minutes of atmospheric exposure. After 30 minutes, gasoline (evaporated) could be identified on the jeans, the cotton t-shirt, and the sweatshirt. After 60 minutes gasoline (evaporated) could be identified only on the jeans. This study demonstrates that in an enclosed room the volatile components of gasoline may adhere to various types of clothing material. However, a positive identification of gasoline by the laboratory would be unlikely even if the clothing was packaged immediately after the exposure. After being exposed to the environment for a short period of time, any indications of gasoline are eliminated. However, when liquid gasoline comes in contact with clothing, the laboratory can make a positive identification of gasoline if the clothing is collected a short period of time after the exposure. Gasoline, Clothing, Ignitable Liquid B121 High Speed High Resolution GC/MS of High Explosives Chin-Chin Lim, MSc, MBA*, Poh Ling Chia, BSc, Sok Hwee Irene Tan, Kim Lian Janice Kuah, MSc, and Ming-Kiong Michael Tay, PhD, MBA, Centre for Forensic Science, 11 Outram Road, Singapore, 169078, Singapore Attendees will learn the use of high speed GC/MS to perform rapid, efficient, and reliable analysis and identification of trace levels of high explosives in post-blast debris. 91 High speed high resolution GC/MS has the potential to be a powerful tool in routine analysis by increasing sample throughput and improving laboratopry productuivity. Significant savings in time and money can be acheived through this technique without sacrficing sepration efficiency. This presentation will impact the forensic community and/or humanity by providing useful for the fast analysis of organic explosive mixtures in postblast investigation where a short “time-to-result” is urgently required. Introduction: The rapid, sensitive, and specific identification of trace levels of high explosives has taken center stage with the escalating threat of terrorist acts worldwide. High speed high resolution GC/MS has the potential to be a powerful tool in explosive analysis by increasing sample throughput (save time and money) and improving laboratory productivity. This paper focuses on a faster separation of a mixture of 14 organic explosives with improved resolution compared to the conventional method. It explores the scope and limitations of the speeding approaches for the analysis of high explosives. The objective is to demonstrate that rapid analyses are possible and can be achieved without sacrificing resolution and separation efficiency. Three columns were used in this study: the conventional long middle bore column (30 m x 0.25 mm x 0.25 µm), a short middle bore column (10 m x 0.25 mm x 0.25 µm), and a short narrow bore column (10 m x 0.1 mm x 0.1 µm). Different speeding approaches were adopted and each of their significance and impact in reducing the analysis time was evaluated by comparing the column efficiency, peak resolution, areas, heights, and widths. The speeding approaches used include: (a) (b) (c) (d) (e) Fast temperature programming Higher initial oven temperature Increased carrier gas velocities Pressure programming Decreased length, bore size, and film thickness of columns The effects of a temperature programmed injection temperature on peak broadening and sample decomposition were also investigated using a programmable temperature vaporiser (PTV). Peak resolution, areas, heights, and widths obtained under PTV conditions were compared to those performed using various constant injection temperatures. Materials and Methods: Instrumentation The analyses were carried out using a GC/MS with programmable temperature vaporiser (PTV). Three columns were used: (a) HP-5MS (30 m x 0.25 mm x 0.25 µm) (b) PTE-5MS (10 m x 0.25 mm x 0.25 µm) (c) DB-5 (10 m x 0.1 mm x 0.1 µm) The analytes were injected into the PTV at isothermal temperature and with temperature programming, using different ramp rates. Analytes: The explosive mixture used in this study comprises of a 70 ppm solution of nitroglycerin (NG), ethylene glycol dinitrate (EGDN), 2-nitrotoluene, 3-nitrotoluene, 4-nitrotoluene, 2,4-dinitrotoluene, 2,6-dinitrotoluene, 2,4,6-trinitrotoluene (TNT), 4-amino-2,6-dinitrotoluene, 2-amino-4,6-dinitrotoluene, nitrobenzene, 1,3-dinitrobenzene, 1,3,5-trinitrobenzene, and cyclotrimethylene trinitramine (RDX). Results and Discussion: Characteristics of analytes The 14-component explosive mixture comprises of analytes which are thermally labile (requiring low injection temperature) and those which are non-volatile with high boiling points (requiring high injection temperatures). Conventional GC/MS Conventional GC/MS for the analysis of high explosives is performed using a long middle bore DB-5MS column (30m x0.25mm x 0.25 µm) at an injector temperature of 150°C and oven temperature of 50 to 200°C at 15°C/min. The analysis time is 14.8 minutes. The objective of this study is to apply various speeding approaches to reduce this long analysis time to achieve a short “time-to-result” analysis. 92 High speed GC/MS (a) Injection system Sample introduction is the most critical step in high speed GC because it affects band broadening. The advantage of a PTV is that it combines a cool injection step with controlled vaporisation. The smaller i.d. liner of the PTV can also be heated or cooled more rapidly and this helps to reduce peak broadening. There was poor sensitivity of the high boiling point analytes at an isothermal injection temperature of 100°C while thermally labile components decomposed at higher temperatures. On using a PTV temperature program from 100 to 250°C, the decomposition of the thermally labile analytes was greatly reduced and better detection limits of the higher boilers were obtained. The ramp rate for the PTV was investigated at the various temperature programming rates from 50°C/min to 250°C/min. At low ramp rates, resolution was poor for two critical pairs. There was a marked improvement in peak separation, peak shape, area, and height at higher ramp rates. Hence, PTV was employed in all the analysis with the three columns. (b) Long middle bore column (30 m x 0.25 mm x 0.25 µm) It was observed that only the initial oven temperature has some impact on reducing analysis time but overall results of the other speeding approaches are relatively insignificant with some loss of resolution. Although increasing the ramp rate produced significant changes in analysis time, peak separation was poor and hence, faster temperature programming rates could not be implemented in this analysis. (c) Short middle bore column (10 m x 0.25 mm x 0.25 µm) An effective way to reduce analysis time is to reduce the column length. Increasing the ramping rate from the conventional 15°C/min to 70°C/min produced the most significant change in analysis time with good resolution. Combining all the speeding approaches resulted in a significant reduction in analysis time by 80%. (d) Short narrow bore column (10 m x 0.1 mm x 0.1µm) Narrow bore columns with thin films are more efficient and have better resolution. It is also possible to work at higher linear gas velocities with less loss in efficiency. Increasing the ramping rate from the conventional 15°C/min to 70°C/min produces very significant changes in analysis time. The shortening of column length, constant pressure programming and higher initial oven temperature produced marginal reductions in analysis. Combining all the speeding approaches resulted in a significant reduction in analysis time by 80%. Comparison of the three columns (a) Analysis time There was a significant 4.5 fold reduction in analysis time for both the short middle and short narrow bore columns but a marginal 1.5 fold reduction was obtained for the long middle bore column. (b) Column efficiency Plate height was used as a measure of the efficiency of the columns. Plate heights obtained for the 3 columns were similar. (c) Peak width and resolution The peak widths were similar for the two short columns, which were only ½ the width of the peaks obtained for the long middle bore column. The resolution-calculations were carried out using the two critical pairs. The two short columns gave good resolution for the two peak pairs and a baseline separation of these pairs was obtained. The data indicate that the gain in speed was achieved without sacrificing resolution. (d) Repeatability There is usually a concern that high speed GC will produce peaks that are not ideal in shape with the presence of fronting and tailing, poor baseline resolution or poor precision of the retention * Presenting Author times and peak areas and heights with faster temperature programming. To determine the precision of the high speed GC method, six repeated injections of the explosive standard mixture were performed. From the results obtained, the repeatability of the retention times, peak areas, and heights in high-speed analyses were found to be satisfactory.%RSD of these parameters for the columns were found to be similar to each other. (e) Limits of detection (LODs) Better LODs were observed for the narrow bore column. This is probably due to the increased signal to noise ratio for the much narrower peaks. (f) Limiting factors The use of high speed GC is usually hindered by a lack of instrumentation. A high power GC oven is required to allow for high programming rates up to 100°C/min. Pressure can also become a limiting factor for a number of speeding approaches. The use of shorter columns is limited by the minimum inlet pressure required for stable operation of the carrier gas system while long narrow bore columns require sufficiently high pressures. The use of columns with a reduced i.d. is one of the most logical approach for high speed GC but sample capacity has to be compromised. Sample capacity of a column with small i.d.s is lower than that of a middle bore column. Sample matrix Sixteen different materials were swabbed with acetone and the swabs were concentrated and analysed using the narrow bore column. The materials tested include: metals, plastics, paper, polymeric materials, wool, soil, sand, tape, and rubber. Only two materials gave co-eluting interference with two of the analytes. A plastic container had a co-eluting peak with TNT and the chromatographic profile of newspaper had a peak that coeluted with EGDN. Conclusion: This new technique allows the Criminalistics Laboratory to perform rapid, efficient, and reliable identification of trace levels of organic explosive residues in post-blast exhibits without sacrificing resolution and separation efficiency. There is a significant decrease in analysis time by 4.5 fold (from 14.8 minutes to 3.3 minutes) Fast temperature programming rates significantly decreased the analysis time, much more than other speeding approaches. Compared to conventional GC techniques, a significantly higher sample throughput is now possible. The new approach has the additional advantages of lower limits of detection, better peak separation, and resolution and a reduction in thermal degradation. High Speed GC/MS, Programmable Temperature Vaporiser, Fast Temperature Ramping B122 Realizing the Science Behind Canine Detection of Explosives Ross J. Harper, MSc*, José R. Almirall, PhD, Douglas Heller, PhD, and Kenneth G. Furton, PhD, International Forensic Research Institute, Florida International University, Department of Chemistry and Biochemistry, Miami, FL 33199 Attendees will learn scientifically sound methods and applications of canine detection of explosives, an understanding of the ability and limitations of canine detection, and the future of this recognised method of explosives detection. This presentation will impact the forensic community and/or humanity by allowing members of the forensic community who are not familiar with canine detection to become familiar with the abilities and limitations of explosives detection canines, whilst at the same time providing specific scientific suggestions to those in the field who wish to improve upon training and operating practices. * Presenting Author Police and private agencies have used canines for many years now, to locate items of forensic interest such as explosives, controlled substances, currency, ignitable liquid residue, hidden or missing persons, and human remains. Up until recently however, this practice has remained more of an art than a true science, and with the increasingly strict standards of forensic evidence admissibility, including Frye and Daubert, validation of canine detection, to provide peer reviewed acceptance of scientifically sound practices, is becoming urgently required. Although viewed as older technology, now competing with microchip laboratories and micro sensors, the canine must still be considered one of the best real-time methods for the detection and location of explosives. Solid Phase Microextraction has been combined with Gas Chromatography - Mass Spectrometry (SPME-GC-MS) and Gas Chromatography - Electron Capture Detection (SPME-GC-ECD) to analyse the headspace of samples of explosive obtained from local and state law enforcement agencies. The SPME-GC-ECD method used in this study is also being optimised for potential field application. A rapid SPME exposure followed by a 15 minute GC program provides ECD spectra of all EPA 8330 explosives, in addition to EGDN, NG, PETN and DMNB. A dual column method is under investigation for confirmatory analysis. Samples analysed include cast explosives, plastic explosives, detonation cords, powder explosives, and commercially available nonexplosives training aids. Results of analysis indicate highly similar odor signatures within the cast and the plastic explosive groups. The cast explosives feature predominantly 2,4-dinitrotoluene and other aromatic mono-, di- and tri-nitrates, whereas the 2,3-dimethyl-dinitrobutane taggant and 2ethyl-1-hexanol plasticiser are common in the plastic explosives. Unlike the high explosives, the odor signatures of the powder explosives are not similar within the group, with the odor chemicals highly dependent upon the manufacturer’s choice, with no common chemical observed within all samples. Chemicals detected within the powder explosive headspace include dinitrotoluenes, ethyl centralite, diphenylamine, nitrodiphenylamine and nitroglycerin. The non-explosive training aids have been analysed in the same manner as the explosive samples, and whilst TNT, RDX and PETN were detected at low levels in the headspace of the silica powder based products, the petrolatum jelly aids do not have significant odor headspaces, above the petrochemical background matrix. Fieldwork with local and state police agencies is currently a multifaceted project. The effectiveness of commercially available training aids has been studied using canines previously trained using actual explosive material, in addition to use as a training medium for new canines. It has been observed that dogs trained on actual explosive have difficulty in locating the non-explosive counterparts, but that dogs trained on the inert aids have little problem in crossing over to the real explosives. Fieldwork is also ongoing with the odor chemicals detected in the headspace of the explosive samples. To date, 2,4-dinitrotolene and 2-ethyl-1-hexanol have been identified as active odors for cast/powder and plastic explosive respectively. The 2,3-dimethyl-dinitrobutane taggant is not utilised by the canines, and is classed as an inactive odor, along with diphenylamine and ethyl centralite, all of which the canines show little or no interest towards. Further research has focussed upon the permeability of explosive odors through assorted plastic and polymer containers. By varying the thickness and chemistry of the polymer, the rate of odor permeation may be controlled. In-house training aids have been developed utilising the active odors and the permeation study, and these aids are currently in applicability trials with selected law enforcement agencies. Work continues to identify the active odors of explosive samples, and to present figures on the reliability of local and state agency trained canines when presented with hidden explosives. Development of inert training aids that mimic explosive odor is ongoing, and these aids have been designed to release odor at a controlled rate of permeation, giving the ability to ‘calibrate’ the canines and calculate minimum detection levels. It can be shown, with the data on reliability, and minimum detection levels, that canine detection may be considered a scientifically sound method of detection. Canine Detection, Explosives, Solid Phase Microextraction 93 B123 Raman Microscopy of Low Explosives and Their Combustion Products Ming-Kiong Michael Tay, PhD, MBA*, Chin-Chin Lim, MSc, MBA*, Poh Ling Chia, BSc, Yuen San Vicky Chow, MSc, and Wanjing Su, Centre for Forensic Science, Health Sciences Authority, 11 Outram Road, Singapore, 169078, Singapore After attending this presentation, attendees will learn the use of Raman microspectrophotometry on trace level detection of common ingredients of low explosives and their combustion products This presentation will impact the forensic community and/or humanity by illustrating the usefulness of Raman for trace level detection of low explosives and their combustion prioducts with minimal sample preparation. Background: Inorganic low explosives usually consist of oxidiserfuel mixtures. They can be made using ordinary laboratory chemicals, ammonium nitrate fertilizer, black powder, and pyrotechnic materials from sparklers, flash powders, and fireworks. Formulations and recipes are readily available on the Internet. Common oxidising agents used are nitrates, perchlorates, and chlorates of sodium, potassium, and ammonium. Fuels include sulphur, finely divided aluminium, magnesium, titanium, zinc, charcoal, sugar, red gum and shellac. This paper illustrates the usefulness of dispersive Raman microspectrophotometry for the detection of low explosives and their combustion products. Major techniques for the analysis of low explosives include stereomicroscopy to mechanically separate individual particles and crystals, scanning electron microscopy with energy dispersive x-ray analysis (SEM/EDX) to determine elemental composition and morphology of particles, polarised light microscopy (PLM) to screen and differentiate crystals, microchemical (spot) tests, ion chromatography (IC) for soluble ions in water extracts, and Fourier Transform infrared (FT-IR) microspectrophotometry for molecular structure elucidation. Current techniques however suffer from several limitations. SEM/EDX is non-destructive and extremely sensitive and powerful for elemental composition but cannot identify molecular and ionic species. Microchemical tests are destructive and often lack sensitivity and specificity; expected color changes may also be masked by interferences. Ion chromatography is a destructive separation technique and although sensitive, is based on matching retention times since conductivity detectors do not discriminate ionic species. In recent years, advances in Raman spectrometer design, improved lasers and detectors, faster acquisition of Raman spectral data, increased sensitivity, easier optical alignment and instrument operation, new approaches to reduce fluorescence masking, and greater affordability have led to the growing popularity of Raman spectrometry as an alternative vibrational spectroscopic technique. Raman spectroscopy has widely been reported as having several advantages over infrared spectroscopy. Raman analysis requires minimal or no sample preparation. Unlike IR spectroscopy, the Raman technique is not affected by atmospheric moisture. Glass cells and aqueous solutions may be used to obtain Raman spectra and the sample can often be examined in situ non-intrusively. The Raman spectrum covers a wider range from 150 to 4000 cm-1, compared to micro-FT-IR which often uses a mercury cadmium telluride (MCT) detector limited to the range 750-4000 cm-1. The far-IR region below 750 cm-1 often has absorption bands with useful molecular information. The IR and Raman spectra of a sample may differ considerably and hence each technique can provide additional, complementary information regarding the sample. Infrared-inactive bands may be observed by Raman spectroscopy, and bands that are weak in IR spectra are often strong in Raman spectra. Materials and Methods: Analyses were performed on a Renishaw RM1000 Raman spectrometer. The spectrometer was calibrated using a silicon standard. A) General comparison of oxidisers and combustion products To ascertain whether oxidisers and their combustion products 94 can be differentiated using Raman microscopy, the following solid phase salts were analysed and their spectra compared: potassium nitrate, potassium chlorate, potassium perchlorate, sodium nitrite, potassium chloride, potassium carbonate, and potassium sulphate. The specificity of Raman spectra for solid phase nitrates, chlorates, and perchlorates were also studied by varying the cation and monitoring Raman shifts for the different vibrational modes. Small amounts of the solids were placed on ordinary glass slides. Samples mounted on graphite stubs for SEM/EDX analysis could also be directly analysed in the Raman microscope. B) Repeatability of Raman shift measurements Repeatability of Raman shifts was studied by performing two runs a day over three successive days. Up to 4 principal peaks were monitored for the six runs. The mean, standard deviation and coefficient of variation were then calculated. This study was applied to 8 oxidisers and 4 combustion products. C) Analysis of aqueous solutions As water is often used to extract post-blast debris for analysis, aqueous solutions of the oxidisers and combustion products were also analysed using the technique. Solutions of 1% and 5% (w/v) of 24 salts prepared using DI water. A sample volume of 15 µL (0.15 - 0.7 mg solute) of solution was drawn into and sealed in a capillary tube (3.5 cm long x 1.5 mm outer diameter.). The tube was placed directly in the path-length of the laser beam. The effects on Raman shifts from hydration of ions dissolved in aqueous solution were investigated. D) Analysis of mixed salts recrystallised from aqueous solutions Ion exchange upon crystallisation of mixed salts from the evaporation of aqueous solutions was studied. A small volume (30 µL) of 1% or 5% solution (containing 0.3-1.5 mg solute) was placed in a cavity slide and evaporated to dryness using the “quick dry” approach or the “slow dry” approach. “Quick dry” consisted of drying on a hotplate at 60oC for 5-10 minutes. “Slow dry” was carried out under natural unforced conditions at ambient humidity and 22oC for 6-8 hours. The crystals were sampled in the same way as for original solid materials. A small amount of the solid was placed on glass slide or mounted on a carbon stub. Results and Discussion: A) General comparison of oxidisers and combustion products The number of peaks, Raman shifts and peak intensities and widths of common oxidisers and combustion products were found to be significantly different, allowing the different oxyanions, sulphate nitrate, chlorate, perchlorate, nitrite, chloride, carbonate, and sulphate to be readily differentiated. Raman spectra are unique for the different functional groups (anions). For each anion, the solid phase Raman spectra were found to be specific for each mating cation. B) Repeatability of measurements Good repeatability was obtained for Raman measurements. The worst case occurred for sodium salts; slight irregularities in peak shapes contributed to the apparent shifts. Since identifications are based on several peaks in the spectrum, this repeatability was sufficient for conclusive identification of oxidisers and their combustion products. C) Analysis of aqueous solutions In aqueous solution, the Raman spectra of nitrates of ammonium, barium, sodium, and potassium could not be differentiated. The effect of cations disappeared for the salts in solution. The spectra showed higher base-line, peak broadening, loss of small peaks and shoulders, shorter peaks, and lower resolution. Shifts were observed in the peaks between solid state and dissolved salts. Raman spectra of recrystallised oxidisers were indistinguishable from those of the original solid salts. * Presenting Author D) Analysis of mixed salts recrystallised from aqueous solutions Ion exchange was observed in some cases but not in others. Crystallisation is influenced by the relative solubilities and ionic radii of the competing ions in solution. Conclusion: Results indicate that Raman microscopy is a very powerful non-destructive technique that can easily identify oxidisers at low detection limits. Spectral resolution was of the order of 1 cm-1, and Raman spectroscopy was found to conclusively identify ionic species, even in aqueous solutions. High spatial resolution was achieved; a microscope objective of 50x gave a laser spot size of 2 microns on the sample, indicating a high sensitivity of approximately 20 picograms of material. Combined with elemental information from SEM/EDX, the new Raman technique allows quick identification of both the cationic and anionic moieties of oxidisers with minimal sample preparation. Raman Microscopy, Low Explosives, Oxidisers B124 Raman Microscopy of Low Explosives Obtained From Sparkler Material Ming-Kiong Michael Tay, PhD, MBA*, Chin-Chin Lim, MSc, MBA*, Poh Ling Chia, BSc, and Wanjing Su, Centre for Forensic Science, Health Sciences Authority, 11 Outram Road, Singapore, 169078, Singapore The goal of this presentation is to present results of a study on the Raman microspectrophotometry of common ingredients in sparklers and their combustion products. This paper is the sequel to an earlier paper presented in this Meeting, titled “Raman microscopy of low explosives and their combustion products.” This presentation will impact the forensic community and/or humanity by illustrataing that sparklers are a convenient source of low explosives and that burnt and unburnt sparkler materials can be easily identified using Raman microscopy. Background: Low explosives are often used in homemade bombs and improvised explosive devices (IEDs). Although low explosives normally deflagrate burning very rapidly at a subsonic propagation velocity, they can generate the gas pressure necessary for an explosion if confined within an appropriate container. Inorganic low explosives usually consist of oxidizer-fuel mixtures. They can be made using ordinary laboratory chemicals, ammonium nitrate fertilizer, black powder, and pyrotechnic materials from sparklers, flash powders, and fireworks. Sparklers are a convenient source of low explosives for small homemade devices. Two main types of sparklers are widely available: wire sparklers and tube sparklers. Wire sparklers are more common and consist of pyrotechnic mixture coated on a length of fairly rigid metal wire. Variants of wire sparklers include dipped sticks where the pyrotechnic mixture is coated on a stick, whistling sparklers that emit a shrill whistle, and crackling sparklers that produce a series of sharp popping or crackling sounds as they burn. Tubed sparklers (or cylindrical fountains) contain the pyrotechnic mixture in loosely-filled cylindrical tubes that resemble pencils in size and shape. A priming mixture is often painted on the tip of sparklers to make ignition easier. Sparklers are stable at room temperature and normally safe to be hand-held for ignition. Lighting the ignition tip results in a flame that propagates down the wire or tube, producing a flame envelope surrounded by shower of colored sparks. Pyrotechnic materials can be easily scraped off wire sparklers or emptied out of a large number of tubed sparklers. Alternatively, wire sparklers can be bundled together using tape to increase explosive effects. The typical pyrotechnic composition of sparklers is a mixture of oxidiser, fuel, combustible binder (eg, sugar, starch, gum arabic and shellac) and a color agent. Fuels include fine metal powder (glazed iron, aluminium, zinc, magnesium, titanium), meal gunpowder, sulfur and charcoal. Oxidisers are usually nitrates, chlorates or perchlorates. In wire sparklers, the iron or mild steel support wires also serve as a heat conductor, promoting the * Presenting Author smooth propagation of the pyrotechnic reaction along the sparkler. Combustion produces a slight pressure, which ejects glowing metal particles, forming sparks that cool quickly. Nature of Study: In the earlier paper titled “Raman microscopy of low explosives and their combustion products,” Raman microscopy was shown to be a rapid, non-destructive and highly specific technique for identifying inorganic low explosives and their combustion products at low detection limits. Raman microscopy identifies both the cationic and anionic moieties of these substances with minimal sample preparation. This second paper illustrates the application of dispersive Raman microscopy to the identification of sparkler materials and their combustion products. Scanning electron microscopy with energy dispersive x-ray analysis (SEM/EDX) is a powerful screening technique to determine elemental composition, particle morphology and the presence of organic constituents, but cannot identify molecular and ionic species and therefore needs to be complemented by a spectroscopic technique such as Raman microscopy. Raman analysis does not give signals for metals, chlorides and some oxides but these elements are readily detected in SEM/EDX analysis. Water extraction allows separation of water-soluble components and oxidisers from the matrix, simplifying subsequent identifications. A twoprong approach combining SEM/EDX and Raman microscopy is recommended for the conclusive identification of inorganic explosive ingredients. Materials and Methods: Analyses were performed on a Renishaw RM1000 dispersive Raman spectrometer. The spectrometer was calibrated using a silicon standard. Materials studied were several commercially available wire sparklers. Sparkler materials were analysed before and after combustion, in solid form and aqueous extracts. Sampling method for Raman analysis: Small amounts of the unburnt, burnt or recrystallised solids were placed on ordinary glass slides. Samples mounted on graphite stubs for SEM/EDX analysis were also directly analysed in the Raman microscope. Aqueous solutions were taken up into glass capillary tubes for direct analysis in the Raman microscope. Analysis of unburnt sparkler tip and body: Materials were carefully scraped and removed from the different layers of pyrotechnic coating for SEM/EDX and Raman analyses. These materials were also extracted with DI water, and the extracts analysed in aqueous solution and as dried recrystallised form. Analysis of combustion products: A sparkler was burnt and materials were carefully sampled from different parts (tip, middle, bottom, exterior, core) of it for SEM/EDX and Raman analyses. DI water was also used to extract post-combustion debris for any residual water-soluble products. Results and Discussion: The results for a 12-inch whistling sparkler are presented in this abstract. Additional results for other sparklers will be presented and discussed further during the oral presentation. The whistling sparklers came in a box of 100, priced at about $4-5 USD a box. Each sparkler was 29.5 cm long and had a pyrotechnic coating measuring 13.7 cm in length. The average weight of a sparkler was 6.05 g. The average amount of pyrotechnic coating on each sparkler was 3.55 g. Each sparkler had a greenish ignition tip, a grey exterior and a white core. The white whistling core was present only in the upper part. Analysis of unburnt whistling sparkler materials: Raman analysis indicated that the unburnt sparkler tip contained mostly barium nitrate (oxidiser) and antimony sulfide (fuel and enhancer of sound effects). The white core of the unburnt sparkler contained mainly potassium perchlorate (oxidiser) and a small amount of organic ingredients. T he exterior contained mainly barium nitrate, carbon and silicon nitride (a refractory material that raises the flame temperature and prevents caking). SEM/EDX indicated the presence of aluminium powder (fuel). Analysis of solids recrystallised from water extracts of unburnt sparkler: Recrystallised materials from water extracts of a mixture of exterior and core materials contained barium nitrate and potassium perchlorate, indicating that no ion exchange occurred during recrystallisation. Analysis of combustion products: The whistling sparkler burned with golden sparks for 45 s and emitted a shrill whistling sound for the initial 5 seconds. The ignition tip and the white whistling core were consumed, 95 leaving a hollow shell in the upper part of the wire and a reduced diameter in the rest of the wire. The average weight of solid residues left was 1.8 g and the loss on ignition was 1.74 g (49% loss). The upper part of the burnt body yielded materials with no significant peaks in Raman analysis, but SEM/EDX analysis indicated barium, aluminium, oxygen potassium and chlorine peaks, consistent with barium oxide, aluminium oxide, and potassium chloride. The lower burnt body also contained carbon with trace amounts of silicon nitride. Material from the burnt body gave little solid residue when extracted with water. Conclusion: Sparklers are a convenient and readily available source of premixed low explosives for making small IEDs. The inorganic ingredients used in sparkler compositions and their corresponding combustion products can be conclusively identified using SEM/EDX and Raman microscopy. Raman microscopy provided rapid, sensitive and unequivocal information on molecular and ionic structures. Metal particles, and some metal oxides and chlorides were not detected by Raman analysis but were easily detected by SEM/EDX analysis. A two-prong approach combining the strengths of Raman microscopy and SEM/EDX offers clear advantages over traditional techniques such as polarised light microscopy and microchemical (spot) tests. The authors are grateful to Mr Wong Soon Meng of the Criminalistics Laboratory for performing SEM/EDX analyses, and taking photographs and scans for the Powerpoint presentation. Raman Microscopy, Sparklers, Low Explosives B125 Identification of Household Chemicals Used in Small Bombs via Analysis of Residual Materials Kirstin McDonald, BA, BS*, and Melvin Shaw, MS, New York Police Department Laboratory, 150-14 Jamaica Avenue, Jamaica, NY 11432 Attendees will obtain information on methodology for the identification of reactants used in a specific type of bottle bomb (also known as chemical or “MacGyver” bombs) by analysis of product residues that result from the mixing of certain household chemicals with commonly available liquids. This presentation will impact the forensic community and/or humanity by providing forensic examiners with information and data useful in identifying chemical bombs made from household products containing halogenated organic compounds; the identification of an original reactant can be used to narrow the search for possible household chemical sources and thus provide an investigative lead. This presentation will also raise awareness of the increasing popularity of these types of bottle bombs and the availability of “recipes” for them on the Internet. Bottle bombs have commonly been made by combining aluminum foil and either muriatic acid (hydrochloric acid) or lye (sodium hydroxide) drain cleaner and water in a plastic bottle, which is then tightly capped and shaken. The resultant build-up in gas pressure causes the bottle to violently burst, and may spray caustic or corrosive liquid. While these devices are often set off as a mere prank, they have been known to cause property damage and/or physical injury. Recently, a growing number of bottle bombs encountered at the New York City Police Laboratory have been not of the afore-mentioned type, but the combination of certain pool and toilet tank tablets with rubbing alcohol. A number of household chemicals contain certain halogenated organic compounds in sufficient concentration which, when mixed with water or other liquids will produce a large volume of gas upon decomposition. These products include pool and spa chlorinators and brominators, and automatic toilet tank cleaners. The first step in this study was to identify products that contain the organic compounds used in these types of devices. This was done by searching stores, browsing online vendors, and looking through the National Institutes of Heath Household Products Database (http://householdproducts.nlm.nih.gov/). 96 In the second step, several pool and toilet chemicals containing such compounds as trichloroisocyanuric acid, sodium dichloroisocyanurate, dichloro-5,5-dimethylhydantoin, and bromochloro-5,5-dimethylhydantoin were mixed under controlled conditions with tap water, rubbing alcohol, and soda-pop. The third step was to take reagent quality compounds in the above list and react them with distilled water and solvent grade isopropyl and ethyl alcohols. The solid residues remaining from each were dried, and then analyzed using gas chromatography-mass spectrometry (GC-MS), Fourier Transform infrared spectroscopy (FTIR), dispersive Raman spectroscopy, and x-ray fluorescence (XRF) and x-ray diffraction (XRD). This multi-instrument approach allowed for several points of comparison between the chemical products of steps two and three and reagent grade standards. It was found that the original solid reactant was easily identified based on characterization of the reaction product. All chromatograms, spectra, and diffraction patterns from the reactions and standards were retained as a reference for future casework. Chemical Bombs, Household Products, Halogenated Organic Compounds B126 Inter-Laboratory Study on Bone Extraction for Mitochondrial DNA Analysis Constance L. Fisher, PhD*, FBI Laboratory, DNAU II, 2501 Investigation Parkway, Quantico, VA 22135 The goal of this presentation is to present the planned round-robin study to the forensic mitochondrial DNA community and encourage the participation of all commercial and government forensic mitochondrial DNA laboratories in the United States and abroad. This presentation will impact the forensic community and/or humanity by increasing awareness of this study and maximize participation among government, commercial mtDNA laboratories in the U.S. and abroad With the implementation of the National Missing Persons DNA Database, the forensic DNA analysis of remains consisting of bone evidence continues to increase. Since the DNA found in bone evidence is frequently limiting and/or degraded, mitochondrial DNA (mtDNA) analysis is often the analysis method of choice. In addition to the government and commercial laboratories currently conducting mtDNA analysis, several additional forensic mtDNA laboratories, including the FBI’s Regional mitochondrial DNA laboratories, New York City Office of the Chief Medical Examiner, and California Department of Justice, are completing internal validation studies and are preparing to begin casework. It is anticipated that a substantial portion of this casework will deal with remains from missing persons. However, the availability of human bones for training purposes is limited. Furthermore, to date, no proficiency test is available using bones as the evidentiary material. The Mitochondrial DNA Subcommittee of the Scientific Working Group on DNA Analysis Methods (SWGDAM) is assembling an inter-laboratory study comparing the extraction methodologies and sequencing results obtained from a single source of bone sample. The study is designed so that similar bones (ie. toe bones) or sections of a long bone (ie. femur) will be obtained from a single donor and distributed to the participating laboratories. Extraction, amplification, and sequencing of the resulting bone DNA will proceed according to the laboratories’ standard protocols. Although full HV1 and HV2 sequencing results is desirable and recommended, partial mtDNA sequencing results from mini-primer sets may be acceptable. In addition to mtDNA HV1 and HV2 sequencing data, autosomal STR, Y STR, as well as mtDNA coding region and non-HV1/HV2 control region SNP data will be evaluated if obtainable from the bone sample. In addition to submitting results of extraction yield and mtDNA sequencing data, participating laboratories will complete a questionnaire * Presenting Author regarding details of their extraction methodologies as well as amplification and sequencing strategies. Comparison of the results and extraction methodologies may highlight differences in methodologies that can be improved to give greater yield of high quality extracted DNA and/or amplified product. In addition, it is expected that this exercise will lend assurance to the field that subtle differences in amplification and sequencing protocols do not lead to differing mtDNA profiles. In order to provide the most benefit to the forensic mtDNA community, all government and commercial forensic laboratories in the U.S. and abroad currently conducting mtDNA testing are encouraged to participate in this study. In addition, laboratories that anticipate conducting mtDNA testing in the near future are also welcome to participate. Contact information as well as a time-frame for participant response, sample distribution, testing, results submission, and study analysis will be discussed. mtDNA, Bone, Inter-Laboratory B127 The Influence of Skeletal Weathering and Bone Type on MtDNA Analysis Lisa M. Misner, MS*, Michigan State University, Forensic Science Program, School of Criminal Justice, 560 Baker Hall, East Lansing, MI 48824; Douglas H. Ubelaker, PhD, Smithsonian Institution, Department of Anthropology, NMNH, MRC 112, Washington, DC 20560; and David R. Foran, PhD, Michigan State University, Forensic Science Program, School of Criminal Justice and Department of Zoology, 560 Baker Hall, East Lansing, MI 48824 After attending this presentation, attendees will learn in what way bone type and its level of weathering, as well as age and sex of the deceased, can be used to help predict the success of mtDNA typing of skeletal material. By using these types of objective comparisons of bone condition and type presented, and individual age and sex, the forensic biologist should be able to better predict the likelihood of successfully amplifying mtDNA from weathered bone, and devise strategies for targeting amplicon length. This presentation will impact the forensic community and/or humanity by increasing the productivity and efficiency of DNA analysis of skeletal material. Forensic investigation of human skeletal remains may require DNA analysis for identification. In many instances such remains are in poor physical condition owing to weathering. This leaves the forensic biologist with the difficulty of predicting how to commence with the analysis, or whether DNA analysis might be successful at all. The goal of this study was to determine if the degradative state of skeletal remains can be used in a predictive manner with regard to mitochondrial DNA (mtDNA) analysis. Further, bone type was examined to establish what role it plays in DNA recovery from weathered bone. Information regarding the sex and estimated age of each individual was also compared to amplification success. Analysis of Variance (ANOVA) was used to test for differences in amplification success among the samples, considering each of these variables. The likelihood of successfully amplifying DNA based on the outer appearance of a bone, as well as the type of bone in question, and what size DNA fragment should be targeted would allow the forensic biologist to begin analysis on skeletal material with the greatest chance of producing results, reducing both the time and resources spent on analysis of aged skeletal remains. In this study, mtDNA was isolated and amplified from a series of skeletons found in an abandoned cemetery in Pittsburgh, Pennsylvania. The cemetery was utilized by Swiss-German immigrants from 1833 to 1861, after which it was eventually built over and subsequently uncovered during an Interstate expansion project in 1987. Over 700 graves were found, and all remains were analyzed anthropologically. The level of weathering was assessed for whole skeletons, and later for individual * Presenting Author bones, to determine if either or both would be beneficial for predicting PCR success and mtDNA quality. Skeletons were graded on a 0 to 5 scale, with 5 being the most weathered, representing bone that crumbled easily. Multiple bone types from skeletons at five out of six stages (1 – 5) were tested genetically. These same sets of bones were regraded on a 1 to 4 scale based on individual bone weathering. Maximum mtDNA amplification size was assessed using primers that produced amplicons of 107 bp, 220 bp 329 bp, and 402 bp. The analysis of this material allowed researchers to determine statistically what influence skeletal and/or bone weathering conditions have on successful mtDNA analysis, as well as what role the type of bone plays. Variables such as the sex and age of the deceased were also included. By using these types of objective comparisons of bone condition and type, and individual age and sex, the forensic biologist should be able to better predict the likelihood of successfully amplifying mtDNA from weathered bone, and devise strategies for targeting amplicon length. This then, would increase the productivity and efficiency of DNA analysis of skeletal material. MtDNA, Bone Degradation, Skeletal Remains B128 Validation of the Roche Linear Array Mitochondrial DNA HVI/HVII Region-Sequence Typing Kit David R. Fisher, BS*, Jason C. Kolowski, MS, Paul Goncharoff, PhD, Mechthild Prinz, PhD, Howard Baum, PhD, and Robert Shaler, PhD, Office of Chief Medical Examiner, 520 First Avenue, New York, NY 10016 After attending this presentation, attendees will learn that the Roche Linear Array Mitochondrial DNA HVI/HVII Region-Sequence Typing Kit is validated for use as a screening method in forensic casework. This presentation will impact the forensic community and/or humanity by demonstrating the use of the Roche Linear Array mtDNA sequence typing kit that will decrease casework turnaround time by quickly screening for exclusions, thereby allowing the analyst to focus on more probative samples. Mitochondrial DNA (mtDNA) typing is a useful tool in the forensic biology laboratory due to the high mtDNA copy number per cell, high degree of polymorphism, and maternal inheritance. MtDNA typing is often successful in yielding profiles in cases where there is an insufficient amount or quality of DNA for nuclear STR DNA testing. While direct DNA sequencing is commonly used to determine mtDNA sequence variation, the Linear Array assay is a more simple and rapid method (e.g. the procedure can be done in about 2 hours versus several days for DNA sequencing) that can be used as a screening technique in mtDNA casework. This presentation will demonstrate that the Roche Linear Array strips are a specific, sensitive, and robust method for the detection of sequence variation in hypervariable regions I and II (HVI/HVII) of the mtDNA control region. During the course of the validation for the new mtDNA laboratory at the NYC Office of Chief Medical Examiner, several different studies were performed on the Linear Array strips to meet the current SWGDAM guidelines for DNA analysis. Sensitivity studies revealed that the most favorable banding and intensity patterns were achieved with approximately 75ng of amplified DNA. Mixture studies revealed that minor components could be seen at a 50:1 ratio, indicating that the array is highly sensitive. Non-probative samples from different tissue types, including, hair, muscle, and bone, were also typed with the Linear Array strips and were found to be in agreement with typing results from direct DNA sequencing. Typing of all laboratory staff was also performed to maintain an in-house database to rule out possible contamination concerns. Lastly, decontamination procedures, such as cleansing gloves in bleach, were employed to minimize the number of failed negative controls. mtDNA, Linear Array, Validation 97 B129 How Well Can Race be Predicted From mtDNA? Mark D. Leney, PhD*, Joint POW/MIA Accounting Command, Central Identification Laboratory, 310 Worchester, Hickam Air Force Base, HI 96853 After attending this presentation, attendees will learn a technique to predict likely self-identified race from an unknown mtDNA profile and to attach statistical weight to such an attribution This presentation will impact the forensic community and/or humanity by allowing explicit calculation of the likely race of an unknown from mtDNA. While not as accurate as nuclear DNA based techniques designed to estimate biological ancestry this will work to provide additional information for a biological profile of a unknown person where nuclear DNA cannot be recovered and could augment nuclear DNA based techniques by flagging the likely race of the mother of an unknown contributor of mixed ancestry. While mitochondrial DNA (mtDNA) in forensic contexts has generally been used to support or refute identification hypotheses, it also contains information that permits inference of the race of an unknown person contributing the mtDNA profile. The mtDNA population database was compiled to provide the forensic community with frequency estimates for mtDNA hypervariable region (mtDNA HVR) profiles observed in evidence-to-reference matches for a specific population of interest. However, the database can also be used to find the frequencies of matching or closely related mtDNA HVR profiles of unknown origin in several different ethnic/racial sub-samples. As the majority of all profiles in the mtDNA population database are themselves unique within the database, it is no surprise that most evidentiary profiles are also observed zero or one times in the database. While such a low frequency is adequate for establishing the relative rarity of the profile in question, it is not informative of probable racial/ethnic origin of the contributor. Nevertheless, as most individuals are only distinguished by a few polymorphisms in the mtDNA HVR from multiple individuals in the database, the pool of closely related individuals is a much richer source of information for estimation of probable ancestry. Attendees will learn how a simple likelihood ratio can be constructed from the probability of observing closely related profiles to an unknown evidence profile in a specific racial or ethnic sub-group and the probability of observing equally related profiles in the rest of the database. Attendees will then observe that by calculating such a ratio for each racial or ethnic sub-group of interest for all possible degrees of profile relatedness it is possible to determine the peak likelihood ratio for racial or ethnic sub-group attribution. This peak represents the balance between idiosyncratic polymorphic differences that are a result of recent within-lineage mutations and older polymorphic differences that are a result of shared common ancestry. The height of the peak indicates the statistical weight of the racial or ethnic attribution. The attendees will also be able to review validation studies conducted on profiles drawn at random from the mtDNA population database and from unrelated but known mtDNA HVR profiles. Neither race nor ethnicity is an absolute biological category but each does contain a biological component. This shared genetic heritage is what allows this technique to work. However, the mtDNA population database is based on self-identified race and ethnicity and thus various other factors act to degrade the power and accuracy of the technique. The main factors are cultural biases in the reporting or recording of single racial or ethnic categories (when many people are actually of mixed ancestry) coupled with the solely matrilineal nature of mtDNA inheritance. These factors will be reviewed and their impact on careful and proper interpretation of results will be discussed. Additionally the applicability of the technique depends on adequate sampling of the populations in question by the database used. The presentation will briefly cover the diversity in the published database, indicating 98 where the database is adequate and where it falls short, giving examples from casework to indicate potential pitfalls in interpretation when a profile from an underrepresented population is analyzed. This technique offers a novel method for placing an explicit statistical value on inference of racial or ethnic group from an mtDNA profile. This could provide a useful lead in building a biological profile of an unknown contributor where only mtDNA was recoverable (as is often the case where conventional nuclear DNA testing fails). Furthermore it could be used in conjunction with the recently developed techniques that estimate biological ancestry from nuclear DNA markers; for example in a person of mixed ancestry, the use of this technique could provide a likely estimate of the self-identified race of the unknown contributor’s mother. At the Joint POW/MIA Accounting Command it is hoped that the technique will assist case resolution for long-term cold cases by highlighting those fragmentary remains that are likely to be unidentified American casualties and those that are likely to derive from indigenous Asian combatants and civilians from the battlefields of Korea and Vietnam. Race, Ancestry, Mitochondrial DNA B130 A Modified Hair Extraction Technique for Mitochondrial DNA Analysis Jason C. Kolowski, MS*, Paul Goncharoff, PhD, Metchild Prinz, PhD, and Robert Shaler, PhD, Department of Forensic Biology, Office of the Chief Medical Examiner, 520 1st Avenue, New York, NY 10016 After attending this presentation, attendees will learn about a modified technique for the processing of evidentiary hairs for use in mitochondrial DNA testing, including, but not limited to, the documentation, handling, washing, and extraction technique employed by the mitochondrial DNA laboratory at the Office of Chief Medical Examiner in New York City. This presentation will impact the forensic community and/or humanity by demonstrating the evidentiary value of hair, whether collected from a crime scene, taken from a body at autopsy, or lifted from evidence in the laboratory, cannot be underestimated. An improved way to process hair evidence for mitochondrial DNA testing will be useful in forensic casework. In the course of implementing a mitochondrial DNA laboratory for the Office of Chief Medical Examiner in New York City, a validation study on hair extraction was performed to ensure the proper handling and processing of evidentiary hairs in forensic casework. One of the goals was to improve the enzymatic digestion technique and to eliminate the high cost and timeconsuming process of manually grinding hairs, which is also a possible source of contamination. To ensure the cleanliness of the hair, a technique of cleaning the hair was developed in which the evidentiary hairs were immobilized on a moist membrane following sonication in a detergent. The membrane, being liquid permeable, allowed the hair to be washed with a variety of reagents and dried in open air without risking the loss of the sample. An enzymatic digestion technique was modified and optimized using regular laboratory-grade reagents, resulting in the complete digestion of a standard 2 cm cutting of a hair shaft in under a half-hour, in nearly all of the samples tested. A variety of studies were carried out on the quality of obtaining viable sequence data from previously mounted hairs, as well as a sensitivity study on the necessary length of the hair prior to enzymatic digestions. Several studies on chemically treated/dyed hair were carried out as well. The enzymatic digestion technique was successful in obtaining mitochondrial DNA from all types of hair, regardless of the mountant or fixative, regardless of chemical alteration or dyes, and from as little as 2 mm of hair shaft. A high success rate was observed in obtaining highquality mitochondrial sequence data from all of the hair extracts (regardless of complete digestion of the hair shaft), initially using Linear Array strips from Roche Applied Science, and confirmed by DNA sequencing and 3100 analysis. * Presenting Author The forensic documentation of the hairs, prior to the processing for extraction, was done on a stereomicroscope from Mideo Systems Inc., allowing for the rapid identification, measurement, and photography of the hair, including the region of interest that would be submitted for the extraction of mitochondrial DNA. Furthermore, the use of dead-air hoods from Labconco and the strict adherence to a conservative decontamination policy in a dedicated pre-amplified room served to reduce the overall effects of contamination and sample loss that are common when dealing with mitochondrial DNA analysis and evidentiary hairs. Mitochondrial DNA, Hair, Extraction B131 A Human Mitochondrial DNA Database Derived From Casework at Mitotyping Technologies Terry Melton, PhD*, and Kimberlyn Nelson, PhD, Mitotyping Technologies, 2565 Park Center Boulevard, Suite 200, State College, PA 16801 After attending this presentation, attendees will know about the diversity and distribution of mitochondrial DNA control region sequences observed in a caseworking forensic mitochondrial DNA laboratory. This presentation will impact the forensic community and/or humanity by demonstrating Forensic mitochondrial DNA analysis is being applied more broadly and frequently. However, forensic mtDNA practitioners rarely present DNA sequence data that they are gathering, and studies on the mtDNA variation present in North America are few. The data presented here will build the scientific understanding of all practitioners, and enhance the application of this type of DNA testing. Between January 1999 and June 2004, Mitotyping Technologies collected 854 human mitochondrial DNA sequences for an internal database. Sequences were complete hypervariable region 1 and hypervariable region 2 nucleotide sequences encompassing positions 1599816400 and 30-407, respectively. A total of at least 783 nucleotides were available for each sequence. Identical sequences within a case (indicating that a match had been obtained between two or more samples within a case) were included only one time in the database. Therefore the samples are assumed to be randomly collected convenience samples derived from both questioned and known samples (blood, bone, hair, saliva, and other tissues) from most of the 50 United States, Canada, and the Caribbean. In most cases, an ethnic/biogeographical identity for the sequence (African/African American, Asian, Caucasian/European, Hispanic, and Native American) could be assigned based on either haplogroup-specific polymorphisms or information submitted with the samples. The sequences were analyzed for sequence diversity, average number of nucleotide differences, haplogroup identity, and length heteroplasmy. These variables were examined at an overall level, within classically defined mtDNA haplogroups, and by state/geographic region. The results were compared to previously published studies on mitochondrial DNA diversity in the continental U.S. and internationally. Overall, 72% (N=614) of the sequences could be classified as historically or biogeographically European while 17% (N=146) could be classified as historically or biogeographically African. Asian-, Hispanic-, Native American-type or unclassifiable sequences comprised the remainder of the sample. The electropherograms of unclassifiable sequences were reexamined for ambiguities or errors and were determined to be correct, indicating that the inability to classify these sequences was likely due to back mutations that have erased haplogroup-specific polymorphisms. Diversity among the Europe-derived sequences was 0.995, with an average number of nucleotide differences of 8.2. Haplogroups H, I, J, K, T, U, V, W, and X were represented in this group, with diversity values ranging from 0.916 (V) to 1.0 (W, X). The average number of nucleotide differences within these European haplogroups ranged from 2.7 (V) to 7.9 (U). Haplogroup H members, the most frequently observed group in all * Presenting Author published studies of European populations to date, comprised 38.4% of the European samples, and had a diversity value of 0.973 and an average number of nucleotide differences of 3.5. The “common” H1 haplotype (263A?G, 315.1insC) was observed at frequencies of 3.7% in the database overall and 5.7% within all European-derived sequences. Among sequences determined to be historically or biogeographically African, diversity was close to 1.0 (0.999), indicating that nearly every sequence was unique in the database. The average number of nucleotide differences among these sequences was 15.1. Haplogroup L1 comprised 28% of the African-origin sequences, haplogroup L2 comprised 20% of these sequences, and haplogroup L3 comprised 36% of these sequences. Individuals from subgroups L1a, L1b, L1c, L2a, L2b, L2c, L2d, L3b, L3d, L3e, and L3f were represented from these three haplogroups. Diversity within the subgroups was uniformly high (above 0.977) with the exception of subgroup L1a (0.917). Mitochondrial DNA, mtDNA Sequence Database, Criminalistics B132 The FBI’s Regional Mitochondrial DNA Laboratory Program Alice R. Isenberg, PhD*, FBI Laboratory, 2501 Investigation Parkway, Quantico, VA 22135 Aattending this presentation, the forensic community will be introduced to the four regional mitochondrial DNA laboratories funded by the FBI to which they may submit cases for analysis at no-cost. This presentation will impact the forensic community and/or humanity by providing a better understanding of the possibilty for increased throughput of mitochondrial DNA casework at no cost through the FBI’s four regional mitochondrial DNA laboratories which will be accepting casework in late 2005. This presentation will familiarize the audience with a new program initiated by the FBI to increase the availability of no-cost mitochondrial DNA analyses to the law enforcement community. The FBI’s fiscal year 2003 budget included $4,000,000 to establish and operate four regional mitochondrial DNA (mtDNA) laboratories. The Congressional authorization requires the FBI Laboratory to provide technical management and quality assurance for regional labs and the regional labs to conduct mtDNA analysis for both forensic casework and missing persons. Competitive procurement began in June of 2003 at a briefing with about 35 state and local crime labs attending or requesting materials. Twelve applications were received in mid-July and cooperative agreements were awarded in September 2003 to the Arizona Department of Public Safety Central Crime Laboratory in Phoenix, AZ, the Connecticut Forensic Science Laboratory (Department of Public Safety) in Meriden, CT, the Minnesota Bureau of Criminal Apprehension Forensic Science Laboratory in St. Paul, MN, and the New Jersey State Police Laboratory in Hamilton, NJ. All four regional mtDNA labs are state-wide, full-service crime labs accredited by ASCLD/LAB in biology and trace evidence and have established forensic nuclear DNA and CODIS/NDIS programs. Under the cooperative agreements, the FBI will train and equip regional laboratories and authorize mtDNA cases to be analyzed according to the Quality Assurance Standards and Scientific Working Group on DNA Analysis Methods (SWGDAM) Guidelines for mtDNA Nucleotide Sequence Interpretation Guidelines. The regional laboratories will provide facilities, employees, and supervision to conduct mtDNA casework, with the FBI reimbursing direct expenses, including salaries and benefits for up to nine employees, space renovation required for mtDNA operations, equipment and maintenance, supplies, training, and travel. When the regional labs are operational, cases will be directed to them and they will be responsible for performing analysis, reporting results, and testifying, if necessary. Fully operational regional labs will have the capacity to examine 120 cases per year, and the regional lab will retain 25% of this capacity for internal allocation. The remaining 75% will be allocated by the FBI for the regional lab’s service 99 area. The current status of the program and procedures for submission of cases will be discussed. The regional mitochondrial DNA program will more than double the nation’s capacity for no-cost forensic mtDNA casework when fully operational in September 2005. Mitochondrial DNA, Regional Laboratories, FBI Laboratory B133 Beyond Laci Peterson: Using DNA Analysis to Identify Missing Persons John C. Tonkyn, PhD*, California Department of Justice/Missing Persons DNA Program, 1001 West Cutting Boulevard, Suite 110, Richmond, CA 94804 The goal of this presentation is to present to the forensic community the development and implementation of a statewide program designed to utilize DNA analysis to solve cases of missing and unidentified persons. The challenges, both technical and administrative, will be described. The need for a nationally coordinated effort will also be discussed. This presentation will impact the forensic community and/or humanity by providing an understanding of the methodology of identifying persons using DNA, but an appreciation for the need for wider implementation of similar programs. The identification of human remains is one of the primary duties of coroners and medical examiners, and is a critical aspect of homicide investigation when the body is not readily identified. In addition to odontology, fingerprints and anthropology, DNA analysis is a highly discriminating tool that is available to identify remains, provided that there is a corresponding reference sample available, either from biologically-related family members or from the missing person themselves (i.e., baby tooth). However, DNA analysis is expensive and body ID is a low priority for most crime labs, so this tool remains vastly underused. The goal of this presentation is to describe the implementation of a statewide program that is successfully providing investigators with the identity of human remains through a concerted effort involving the state crime lab, the state missing person clearinghouse, coroners, medical examiners and local law enforcement. The California Missing Persons DNA Program was created in 2001 with the express purpose of using DNA databases of both unidentified human remains and reference samples for missing person (either swabs from relatives or items from the missing person) to make identifications. Model collection kits have been devised for the collection of oral swabs from family members of missing persons, and for the collection of human remains. Training, in the form of instructional videos and written publications, has been developed. Outreach has involved the identification of “high risk” missing person cases and unidentified person cases reported to the state clearinghouse, and coordination with coroners and law enforcement investigators for the collection of appropriate samples. Short Tandem Repeat DNA profiles, as well as mitochondrial DNA sequencing data are entered into local databases as well as the new “CODISmp,” which has been developed specifically for use with missing and unidentified person DNA profiles. As most analyses involve comparison between remains and a relative of the missing person, kinship analysis is required for comparison of nuclear DNA profiles. While all states participate in CODIS, the primary categories of samples entered are convicted offender and forensic unknown. Few labs routinely enter profiles from missing or unidentified persons, and fewer still have dedicated resources to this task. It is one of the goals of this presentation to provide a model for other states to consider when implementing a program for the purpose of identifying human remains. Because of the movement of individuals and families between states, a certain percentage of cases either will not be analyzed or compared after analysis. A national program of all states entering both STR and mitochondrial DNA into NDIS is required, similar to the highly successful convicted offender/forensic unknown model. 100 Several dozen identifications have been made to date. Technical improvements in typing human remains will be discussed. These include the following: 1) Improved methods for DNA extraction, 2) qPCR methods that simultaneously quantify the amount of both nuclear and mitochondrial DNA, and 3) the use of a duplex PCR reaction for the simultaneous amplification of both HV1 and HV2 regions of mitochondrial DNA (thereby conserving sample which contains little DNA to begin with). Finally, several successful cases will be described that highlight the issues and technologies describe above. Missing Person, Mitochondrial DNA, Unidentified Person B134 Optimization of Mini Plex Primer Sets for STR Testing of DNA Extracts From Bone Samples 10 – 13 Years Postmortem Jon Davoren, MS*, International Commission on Missing Persons, Alipasina 45a, Sarajevo, 71000, Boania and Herzegovina; Michael Coble, PhD, National Institute of Standards and Technology, 100 Bureau Drive, Mail Stop 8311, Gaithersburg, MD 20899-8311; and Tony Donlon, BS, Ana Milos, BS, and Rijad Konjhodzic, BS, International Commission on Missing Persons, Alipasina 45a, Sarajevo, 71000, Bosnia and Herzegovina The goal of this presentation is to describe some of the PCR optimizations that have been performed on a number of reduced amplicon primer sets for amplification of DNA from 10 – 13 year postmortem skeletal remains. The work presented here will be useful for anyone performing DNA STR testing of bone sample. The goal of this presentation is to describe some of the PCR optimizations that have been performed on a number of reduced amplicon primer sets for amplification of DNA from 10 – 13 year postmortem skeletal remains. The International Commission on Missing persons has developed a DNA led identification system for identification of victims of the conflicts resulting during the breakup of the former Yugoslavia. As a first step in the identification effort bone samples were taken from whole skeletons and tested using a silica-based extraction method and the Promega PowerPlex® 16 system. With the PP16 system results were obtained in approximately 85% of the samples tested. The 15% that were considered failures, during the first phase, had between 0 and 11 loci amplified with most of the failed loci being the longer PCR products. To better understand why these samples were failing STR analysis extracts were assessed using the Applied Biosystems Quantifiler™ Human DNA quantification system. Most of the samples displayed relatively low amounts of DNA (<10 pg/µl) and/or the presence of PCR inhibitors. The results of both the quantification as well as the allelic drop outs in the longer loci indicate that the DNA in these extracts may be degraded to the point where commercially available STR kits are not effective. In an attempt to get more STR data out of the DNA extracts, that failed the initial phase of this testing, the possibility of using reduced amplicon primer sets has been investigated. Initially a concordance study was performed to ensure that the alleles amplified by the new primer sets would reflect the same alleles designated by the PP16 system and the Applied Biosystems AMPFlSTR® SeFiler™ kits. Reduced amplicon primer sets were also optimized for amplification of DNA isolated from a number of bone samples. The optimizations tested include modifications to the PCR buffer, Mg++, and different taq enzymes. The results of the optimizations will be presented and compared to optimizations of the PP16 and SeFiler kits. Additionally the effectiveness of the mini primer sets for STR testing of DNA extracts from bone samples will be discussed in detail. STR, DNA, Mini Plex * Presenting Author B135 A Duplex Real-Time qPCR Assay for the Quantification of Human Nuclear and Mitochondrial DNA in Forensic Samples: Implications for Quantifying DNA in Degraded Samples Mark D. Timken, PhD*, Katie Swango, PhD, Cristian Orrego, PhD, and Martin R. Buoncristiani, MPH, California Department of Justice, Jan Bashinski DNA Laboratory, 1001 West Cutting Boulevard, Suite 110, Richmond, CA 94804 After attending this presentation, attendees will learn the advantages of using a new duplex qPCR protocol for quantifying nuclear and mitochondrial DNA in forensic sample; the implications of using qPCR versus slot blot quantification methods for successfully genotyping degraded DNA. This presentation will impact the forensic community and/or humanity by providing a new tool for DNA quantification in forensic samples; to describe quantification differences, especially for samples containing degraded DNA that can be expected when moving from slot blot to qPCR quantitation methods. This report describes selected results from validation of a duplex realtime qPCR assay that has been developed for the quantification of human nuclear and mitochondrial DNA in forensic samples. This assay was designed to be of general utility for forensic DNA quantifications, but to be particularly useful for the post-extraction analysis of samples that contain degraded DNA. Such samples, though not uncommon in standard casework evidence, are often encountered in evidences from mass disasters, mass graves, and missing persons’ cases. Presently, the initial decision as to how to proceed with analysis of such samples, either by nuclear STR or mitochondrial typing, is commonly based on a slot blot quantification approach that (1) has been reported to underestimate the quantity of nuclear DNA in degraded samples and in samples that contain high levels of microbial contamination, (2) provides no information about the quality (fragment length) of the quantified nuclear DNA, and (3) provides no direct information about the quantity of human mitochondrial DNA in the sample. Due to these quantification deficiencies, the actual forensic analysis of challenging samples often proceeds first by obtaining inadequate STR typing results, and then by using any remaining extracted DNA in an attempt to obtain mitochondrial typing results, the latter analysis typically attempted without any direct knowledge of the presence (or absence) of human mitochondrial genome in the DNA extract. The efficiency and quality of this analysis procedure can be improved substantially by obtaining reliable estimates of the amounts of human nuclear and mitochondrial DNA in these samples. Based on such estimates, an optimal analytical approach can be selected at the outset, leading directly to optimal genotyping or haplotyping results and to a concomitant savings in time, in labor, in reagent/kit costs and of extracted DNA. In the duplex nuclear-mitochondrial qPCR assay developed, the authors quantify a nuclear target sequence that spans the repeat region of the primate-specific TH01 STR locus, a locus that has been used widely for forensic applications. This target sequence is of direct interest for quantification, considering that a primary reason for quantifying human nuclear DNA in forensic samples is to determine the amount of extract to amplify in a commercial multiplex STR PCR kit. For degraded samples, quantification of the relatively long TH01 target (~170-190bp) leads to improved STR typing results, compared to typing results based on quantification by slot blot hybridization. For the mitochondrial portion of the duplex qPCR assay, the authors quantify a short target (69bp) in the mitochondrial ND1 gene. This selection provides a sensitive means for determining the presence of human mitochondrial DNA, degraded or not, in forensic samples. Although the presentation will focus mainly on quantification and analysis of samples containing degraded DNA, the authors will also briefly describe other results from validation work, including studies of precision, * Presenting Author reproducibility, sensitivity, species specificity, and applications to casework-type samples. qPCR, Duplex, Degraded DNA B136 Significance of Detecting Foreign Sources of DNA Under Fingernails Melinda Matte, BSc*, Elida Kafarowski, MSc, Roger Frappier, MSc, and Jonathan Newman, BSc, Centre of Forensic Sciences, 25 Grosvenor Street, 4th Floor - Biology Section, Toronto, Ontario M7A 2G8, Canada The goal of this presentation is to provide the forensic community with a better foundation on which to base opinions with respect to DNA fingernail evidence in casework. This presentation will impact the forensic community and/or humanity by providing the forensic community with data to validate their opinion regarding DNA evidence from fingernail samples in courts of law. Scientists at the Centre of Forensic Sciences (CFS) routinely examine DNA results from fingernail scrapings and clippings in casework. Fingernails are useful in forensic science as they can often be used to generate a DNA profile from the donor and any additional sources. Foreign DNA beneath the fingernails can arise due to the transfer of skin cells or body fluids as a result of intimate physical contact. While the ability to detect foreign DNA profiles from fingernails has been demonstrated, their evidentiary significance is afforded little attention in the literature. Fingernail related items are submitted to the CFS for DNA testing in one of two ways: scrapings or clippings. If the debris underneath the fingernails has been scraped with a wooden applicator stick prior to submission to the laboratory, then all of the scraped debris and the end of the stick are subjected to DNA analysis. In cases where clippings have been provided, the undersides of the fingernails are swabbed at the laboratory prior to DNA analysis. Special attention is given to fingernail clippings with blood staining; for these samples the blood, usually originating from the fingernail donor, is swabbed separately from the remainder of the underside of the fingernails. Experience is that this approach aids in the detection of foreign DNA by separating out samples likely to contain very high amounts of the donor’s DNA. A study performed at the CFS, involving staff of the laboratory and students from a local University, was undertaken to determine the prevalence of foreign DNA underneath the fingernails of individuals who share work or household environments. A total of 39 participants provided 78 samples (from the left and right hands) for DNA analysis by scraping under their fingernails with a wooden applicator stick. Of these samples, only 12% yielded a second source of DNA. When these were further categorized into samples where a significant foreign DNA profile was detected (DNA results for at least 4 STR loci) then the number dropped to only 4%. Further investigation determined that the foreign source of DNA could not be attributed to fellow members of the household or work environment, unless intimate physical contact had occurred. These results suggested that casual contact between individuals is not likely to result in the deposition of a second source of DNA beneath the fingernails. The CFS maintains a database to track DNA results from fingernail related items in casework. Since February 2003 the data from 95 cases have been analyzed (a total of 187 samples). Homicides (67%) and sexual assaults (26%) comprised the majority of cases with fingernail related items. The majority (76%) of samples were submitted as clippings, while 22% were submitted as fingernail scrapings, and 2% contained both clippings and scrapings. Of the samples from fingernail clippings, 34% yielded a foreign source of DNA, while 58% contained only DNA from the fingernail donor, and 8% yielded insufficient or no DNA for analysis. Of the samples of fingernail scrapings, 29% yielded a foreign source of DNA, 40% contained only the fingernail donor’s DNA, and 31% yielded insufficient or no DNA for analysis. 101 The results indicate that the likelihood of detecting a foreign source of DNA on fingernail clippings is greater than in scrapings. This may be a result of the CFS method of sampling any blood separately on clippings to prevent the “swamping out” of a foreign source of DNA with the donor’s DNA during PCR. The data also suggest that fingernail samples can yield useful investigative information when the case history warrants their examination. However in casework, fingernail evidence is only examined if the case history suggests an expectation of finding a foreign source of DNA. Therefore one would expect a much higher prevalence of foreign DNA relative to the controlled study. Additional experimental data regarding both the prevalence and persistence of foreign DNA profiles beneath fingernails under a variety of conditions will be presented. It is expected that this data and research will provide the forensic community with a strong foundation on which to base opinion evidence in courts of law pertaining to the relevance of fingernail findings in casework. Fingernail Clippings, Fingernail Scrapings, DNA B137 Development of a Human-Specific Real Time PCR Assay for Simultaneous Quantitation of Total Genomic and Male DNA Katie M. Horsman, MS*, University of Virginia, Department of Chemistry, McCormick Road, Charlottesville, VA 22901; Jeffrey A. Hickey, MS, Margaret A. Terrill, MSFS, and Robin W. Cotton, PhD, Orchid Cellmark, 20271 Goldenrod Lane, Germantown, MD 20876; James P. Landers, PhD, University of Virginia, Departments of Chemistry and Pathology, McCormick Road, Charlottesville, VA 22901; and Lewis O. Maddox, PhD, Orchid Cellmark, 20271 Goldenrod Lane, Germantown, MD 20876 The goal of this presentation is to describe the development and validation of a duplex real time PCR method for simultaneous determination of total genomic and male DNA concentrations in a forensic sample. This presentation will impact the forensic community and/or humanity by demonstrating the real-time PCR assay which was developed for simultaneous quantitation of total genomic and male DNA should be utilized in forensic DNA analysis as a replacement to slot-blot quantitation. Increased sensitivity, decreased analysis time, specific quantitation of male and total genomic DNA make the assay superior to the slot-blot assay. In addition, the multiplex assay results in time and cost savings when compared to the ABI Quantifiler kits. Historically, the slot-blot method has been used to quantitate human DNA in forensic samples. However, the slot blot method is labor intensive, subjective in interpretation, and not highly amenable to automation, which has led to the development of new methods for quantitating human DNA. Current research1,2 has focused on the use of Real-Time PCR as a more accurate, more sensitive, and less labor intensive method. Real time PCR has the capability of absolute quantitation based upon measurement of the increase in fluorescence signal with each cycle and comparison to a standard curve. A number of detection chemistries are currently available including SYBR® Green detection, fluorogenic probes, and molecular beacon technology. Fluorogenic probe chemistry was utilized in this study, with spectrally distinguishable reporter dyes for each target sequence. With the growing capabilities for discrimination based upon Y-chromosome STR typing, the need to quantitate the male contribution to a sample is becoming increasingly evident. Commercial kits (Applied Biosystems) for human DNA and Y-chromosome DNA quantitation with real-time quantitative PCR are available in singleplex reactions. However, multiplex PCR has the advantage of consuming half of the DNA as compared to the commercial kit in addition to reducing the cost of the analysis. Therefore, a multiplex real time PCR assay has been developed for simultaneous quantitation of both the total human genomic DNA concen102 tration as well as the male DNA component. Primer/probe sets were designed using the Primer Express® (Applied Biosystems) software. For total human genomic DNA quantitation, the amplicon is a 63bp fragment of the TPOX locus. The Y-chromosome amplicon is a 70bp fragment of the sex-determining region (SRY) of the Y-chromosome. Both primer sequences were determined to be human (or higher-primate) specific with a BLAST search as well as tested experimentally with common DNA sources including yeast, E Coli, and mouse. The lower limit of quantitation was determined to be approximately 10 pg for total genomic DNA and approximately 20 pg for male DNA. The upper limit of the quantitation was set at 50 ng, which is appropriate for typical casework samples. This linear range for the real time PCR quantitation is significantly larger than that of the 125 pg to 10 ng range of the slot-blot assay. In direct comparison to a modified QuantiBlot® (Applied Biosystems) assay followed by BioImage® analysis, the average relative error of the multiplex assay was comparable to the slot-blot assay. The average percent error was 11.6% for total genomic DNA and 12.1% for male DNA, compared to 12.8% for the total genomic DNA quantitation by slot blot. The slot-blot analysis is unable to specifically detect the concentration of male DNA in a mixture. A mixture-challenge study indicated the quantitation of 25 pg male DNA was accurate in mixtures of up to 1:5000 male:female DNA. Additional validation experiments included optimization of Mg2+ concentration, comparison of quantitation accuracy to the Quantifiler and Quantifiler Y kits, and assay reproducibility. References: 1. Nicklas JA, Buel E. Development of an ALU-based , Real-Time PCR Method for Quantitation of Human DNA in Forensic Samples. J Forensic Sci. 2003; 48(5): 935-44. 2. Richard ML, Frappier RH, Newman JC. Developmental Validation of a Real-Time Quantitative PCR Assay for Automated Quantification of Human DNA. J Forensic Sci. 2003; 48(5): 1041-6. DNA Quantitation, Real-Time PCR, Slot Blot B138 Application of Real Time qPCR Multiplex Results to Downstream Decision Making in Samples Containing Mixtures of Male and Female DNA Jeffrey A. Hickey, MS*, Orchid Cellmark, 20271 Goldenrod Lane, Germantown, MD 20876; Katie M. Horsman, MS, University of Virginia, Department of Chemistry, Charlottesville, VA 22901; and Lewis O. Maddox, PhD, and Robin W. Cotton, PhD, Orchid Cellmark, 20271 Goldenrod Lane, Germantown, MD 20876 The goal of this presentation is to describe the results of Real Time PCR quantitation of male/female mixtures and the effect of this data on decision making at the point of PCR amplification for STR loci. This presentation will impact the forensic community and/or humanity by providing data from a real time qPCR multiplex for simultaneous quantitation of total genomic and male DNA which can be used to improve amplification decisions. This will assist analysts in determining which DNA samples are likely to provide useful information when amplified for autosomal STR and/or YSTR loci. The availability of real time PCR (qPCR) methods capable of determining total genomic and male DNA in forensic samples has changed and improved the decision making process for STR testing. Other quantitation methods, such as the slot blot, do not provide information regarding the relative amounts of male versus total genomic DNA and are much less sensitive than real time PCR. Knowledge of the male DNA concentration can be used to determine if the sample should be amplified with autosomal and/or Y specific STR loci. In this study, a real time qPCR multiplex capable of detecting male and total genomic DNA in one reaction was used. This assay utilizes * Presenting Author Taqman probe chemistry and contains a primer/probe set for the TPOX locus and a primer/probe set for the sex-determining region of the Y chromosome. The information gleaned from validation of this real-time multiplex was used in concert with validation data from ABI Profiler Plus™, COfiler™, and Promega PowerPlex Y® testing systems to make decisions regarding the most effective approach for sample analysis. Experiments were designed with the purpose of determining if real time results are reliable predictors of both Y STR and autosomal STR amplification results. For this study, several dilution series of male/female mixtures were quantified for total genomic and male DNA using the real time multiplex. After quantitation, all of the samples were amplified using the PowerPlex Y kit and a subset was amplified using the Profiler Plus kit as well. The PowerPlex Y® amplifications were based on the male specific qPCR results, whereas the Profiler Plus amplifications were based on the total genomic DNA qPCR results. The samples were then analyzed using an ABI Prism 310 Genetic Analyzer. The results showed that the real time qPCR multiplex used in these experiments provides valuable quantitation data for both total genomic and male DNA in mixed samples. The real time qPCR results for the male/female dilution series confirmed that this assay detects male DNA in a wide variety of male/female DNA mixtures. The lower range of detection is approximately 12.5pg of male DNA in a 1:5000 male:female mixture. For both PowerPlex Y and Profiler Plus™, the amplifications were successful using the real time qPCR data to calculate the volume to be added to the PCR reactions. When run on the ABI Prism 310, the amplifications resulted in peaks heights within the laboratory’s analysis parameters. Additionally, the real time data proved to be a good indicator of the ratio of male to female DNA. The Profiler Plus peak height ratios for the male/female mixtures were reasonably consistent with the male to total DNA ratios calculated from the qPCR data. These results were also similar to the known male/female ratios in the actual dilutions. From these data and data from the previous validations of Profiler Plus™, COfiler™, and PowerPlex Y, it is possible to draw important conclusions regarding mixed samples prior to STR amplification. Because this assay can accurately detect male DNA in the presence of overwhelming amounts of female DNA, it is possible to decide how much sample to use for PowerPlex Y amplifications. In addition, since the assay can reasonably determine the ratio of male to female DNA in a mixed sample, it is possible to reasonably deduce which samples will result in a useful secondary male profile when amplified for Profiler Plus™. DNA Quantitation, Real-Time PCR, Y STRs B139 Development of Human and Human Male DNA Quantitation Systems Using a Novel, Fluorescent, Two-Primer Real-Time PCR Method Benjamin E. Krenke, MS*, Susan Frackman, PhD, Katharine Hoffmann, BS, Gary Madsen, PhD, Cynthia J. Sprecher, BS, and Douglas Storts, PhD, Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711 The goal of this presentation is to present information on a new approach to DNA quantitation. This presentation will impact the forensic community and/or humanity by demonstrating a new commercial alternative for real-time PCR. Heavily multiplexed Short Tandem Repeat (STR) analysis has become the dominant technology in DNA-based human identification. Although highly informative, these assays require a defined range of template quantity to produce optimal results. Additionally, resources can be conserved with accurate assessment of DNA quality and assessment of minimum quantity. Currently, many practitioners observe either high levels of false negative results (due to lack of sensitivity) or subjective conclusions (due * Presenting Author to visual comparison of band intensities) based on common hybridizationbased methodologies. Amplification-based methods for quantitation provide a high level of sensitivity while real-time methods can deliver a dynamic range that often exceeds end point assays. A numerical output also increases the objectivity of the data interpretation. A real-time PCR method has been developed for the quantitation of total human and human male DNA in purified samples using the specificity of interaction between two modified nucleotides to achieve quantitative PCR analysis. One of the PCR primers includes a modified nucleotide (iso-dC) adjacent to a fluorescent label on the 5´ end. The second PCR primer is unlabeled. The reaction mix includes deoxynucleotides and iso-dGTP, which has been modified to include dabcyl quencher. The only nucleotide incorporated at the position complimentary to iso-dCTP is dabcyl iso-dGTP. The incorporation of the dabcyl iso-dGTP adjacent to the fluorescent dye results in a reduction in signal that allows quantitation during amplification. Associated analysis software has been developed to visualize amplification data from various instrument platforms, plot standard curves and calculate DNA concentrations of unknowns. Relative to other real-time approaches, this methodology provides specificity through the use of fluorescently-labeled primers compared to DNA binding dyes and simplicity compared to probebased quantitative PCR approaches. Data will be presented demonstrating the performance of assays using human autosomal (total human) and human Y-chromosome (male human) targets for quantitation. Forensic Science, DNA Quantitation, Real-Time PCR B140 Utility of Quantifiler™ Y in Resolving Difficult Forensic Cases Huma Nasir, BS*, Gina Pineda, MS, Brekeitrea Minyon Jones, MS, Elaine Schneida, BS, Sarah Corrigan, MS, Sudhir K. Sinha, PhD, and Jaiprakash G. Shewale, PhD, ReliaGene Technologies, Inc., 5525 Mounes Street, Suite 101, New Orleans, LA 70123 The goal of this presentation is to demonstrate to the forensic community the usefulness of Quantifiler™ Y in forensic casework. This presentation will impact the forensic community and/or humanity by demonstrating the usefulness of Quantifiler™ Y in forensic casework. Quantitation of human male DNA and obtaining interpretable male profile are major obstacles during the analysis of a mixture sample containing male and female DNA. Availability of Quantifiler™ Y for quantitation of human male DNA and Y-STR multiplex systems for amplification of short tandem repeat loci on the Y-chromosome (Y-STRs) enables one to overcome these obstacles. It is possible to quantitate male DNA and obtain an exclusive profile of male DNA in a sample containing mixtures of male and female DNA using Quantifiler™ Y and Y-STR multiplex systems, respectively. Scientific Working Group on DNA Analysis Methods (SWGDAM) has identified a set of eleven loci namely DYS19, DYS385a/b, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS438, and DYS439 for forensic analysis. Quantifiler™ Y human male DNA quantitation kit was validated for forensic DNA analysis using the 7500 Sequence Detection System. Analysis for Y-STRs was performed using the Y-PLEX™ 12 system. A combination of Quantifiler™ Y and Y-PLEX™ 12 was used to obtain male profiles in the analysis of 10 forensic cases. Sensitivity of the Quantifiler™ Y on the 7500 Sequence Detection System in the laboratory was 44.5 pg of human male DNA. Quantitation of male DNA by Quantifiler™ Y is based on real time PCR and hence provides scalable information about the quantifiable human male DNA. This approach enables one to obtain male profile from evidence samples. Of the 15 cases investigated, 4 evidence samples provided complete profiles, 2 evidence samples provided partial profiles, and 4 evidence samples provided inconclusive results. Data and strategies used for amplification of evidence samples from these difficult forensic cases will be presented. Y, Y-PLEX 12, Forensic Casework Quantifiler 103 B141 We’re Not Gonna Take It! Girls Fight Back Against Attackers (Case Studies) Noelle J. Umback, PhD*, Louis M. Serico, BS, Ana T. Fernandez, BS, and Mary Quigg, BS, New York City Office of Chief Medical Examiner, Department of Forensic Biology, 520 First Avenue, New York, NY 10016 The goal of this presentation is to present several interesting case studies where in each case, DNA from the assailant was obtained after the victim fought back. To date, CODIS has solved two of these cases. This presentation will impact the forensic community and/or humanity by demonstrating that semen on body swabs need not be the only target for DNA analysis in rape cases. Women who defend themselves during an attack help forensic laboratories by giving them more chances to obtain male profiles from the available evidence. Several recent male-on-female assault cases received for DNA analysis by the New York City OCME Forensic Biology Department have generated CODIS-eligible profiles. Three are from blood, one is from skin cells, and one is from semen. These were all “stranger” rape or attempted rape cases, in different NYC boroughs. The first case of the “pattern” was a home-invasion in which a stranger entered the bedroom of a middle-aged woman during the night, and chased her around her room several minutes in her nightgown. The sexual assault kit was negative, as expected because the attacker never got the chance to complete the rape. She managed to hit him over the head with a chair, drawing blood. Stains on the chest area of her nightgown matched a crime-scene sample collected from the bedroom. The blood proved to be male and was uploaded into CODIS. To date, no hits have surfaced. Another home invader got more than he bargained for when his rape victim managed to get a knife and stab him. No sexual assault kit was collected. There was one semen stain on a washcloth, which yielded no DNA. There were bloodstains on several items from in and around the home, most of which matched the victim. However, drops of blood collected from a nearby sidewalk gave a clean male profile, which was uploaded into CODIS. It hit a convicted offender immediately, making Special Victims Unit detectives very happy, as this crime was one of a string (although the first with DNA evidence) of violent attacks. The suspect was apprehended within 24 hours. A teenage girl was grabbed from behind in the elevator of her own building by a man who told her he had a gun, and pulled her pants down. The attack was recorded by a security camera but the picture was too blurry to be of much assistance in identifying the man. During the struggle in which she was injured as well, she managed to pull the much-larger perpetrator down the stairs with her, knocking off his eyeglasses; and eventually escape into her apartment. Male DNA from the eyeglasses, as well as bloodstains in the case, gave a full CODIS profile; to date this case is unsolved. A woman walking home late on a winter night was grabbed from behind and dragged into a nearby park by a man with a gun. Following completion of the forced sexual activity, the victim kicked him in the groin and sprayed him with pepper spray. Detectives collected a semen sample from the scene which yielded a full CODIS profile. This case has not yet been solved. Last but not least, a woman fended off a rapist in Central Park by twisting the attacker’s penis in her hand when he put it near her face. Meanwhile her dog bit him hard enough to draw blood, which left a stain on her clothing. The male DNA from this bloodstain hit a convicted offender sample in CODIS in the next search after its upload. At the time he was free on parole. Self-Defense, CODIS, Sexual Assault B142 Developmental Validation of a New 17-Y-STR Multiplex System: The AmpFlSTR® Yfiler™ PCR Amplification Kit Lori K. Hennessy, PhD*, Julio J. Mulero, PhD, Chien-Wei Chang, PhD, Robert L. Green, BS, Yixin Li, PhD, Rixun Fang, PhD, Lisa Calandro, MPH, and Cherisse Boland, BS, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404 After attending this presentation, attendees will learn about the validation of a new Y-STR muliplex PCR assay. This presentation will impact the forensic community and/or humanity by providing a new tool available for DNA testing of the Y chromosome. The goal of this presentation is to summarize the results of the developmental validations studies for the AmpFlSTR Yfiler PCR Amplification Kit. This kit was developed in accordance with the guidelines of the forensic community, as defined by SWGDAM and the DNAAdvisory Board’s (DAB) Quality Assurance Standards. Analysis of DNA sequence variation on the Y chromosome has become an increasingly important method in forensic casework, especially in sexual assault samples where there is a relatively small amount of male DNA and a large amount of female DNA. A multiplex PCR amplification system targeting Y-specific loci can simplify interpretation of complex male/female mixtures and yield a high degree of confidence that only the male contributor(s) is being analyzed. The authors have developed and validated a 17-locus Y-STR multiplex system that has a higher discriminatory capacity than the European minimal haplotype. The Y-STR multiplex includes all the loci in the “European minimal haplotypes;” DYS19, DYS385, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, the SWGDAM recommended Y-STR loci; DYS438, DYS439, and six highly polymorphic loci; DYS437, DYS448, DYS456, DYS458, DYS635 (Y GATA C4), and Y GATA H4. By incorporating the automated 5-dye DNA fragment analysis technology and nonnucleotide linkers small amplicon sizes (100bp-326bp) can be maintained and ensure no overlap between allele ranges. 6-FAM™ labeled primers detects DYS456, DYS389I/II, and DYS390. VIC® labeled primers detects DYS458, DYS19, and DYS385 a/b. NED™ labeled primers detect DYS393, DYS391, DYS439, DYS635, (Y GATA C4), and DYS392. PET® labeled primers detect Y GATA H4, DYS437, DYS438, and DYS448. The fifth dye, LIZ® is used to label the GeneScan-500 Size Standard. All loci are coamplified simultaneously in a single tube and analyzed in a single capillary injection. The Yfiler™ kit includes an extensive allelic ladder containing the most common 137 variants observed at each locus. To test the sensitivity of the multiplex male DNA was serially diluted from 4 ng to 31 pg. Complete male profiles above a 50-rfu threshold were reproducibly obtained at 125 pg. Partial to complete profiles could also be obtained at 62.5 and 31 pg. Testing a panel of domestic and farm animals, bacterial species, and female DNA, the kit demonstrated primate and male specificity. No cross reactivity was seen with the various animal or bacterial species and no reproducible peaks above 50-rfu were detected in the presence of 500 ng of female DNA. Complete male profiles were reproducibly obtained at 1000:1 female to male mixtures and the minor contributor was reproducibly distinguishable in male-to-male mixture ratios of up to 10:1. Haplotype concordance was 100% when tested with the NIST standard SRM#2395 and 600 population samples. A database for this new 17 Y-STR multiplex has been created for determining haplotype frequency using population samples from North American, European and Asian population groups. The data are accessible via the internet for haplotype searches. Results demonstrate that the AmpFlSTR Yfiler PCR Amplification Kit is a sensitive, valid, and robust multiplex system for Y chromosome STR analysis. The system can be used in conjunction with Applied Biosystems Thermal Cyclers, and ABI PRISM Genetic Analyzers, and provides the forensic scientist with a complete set of tools for Y chromosome analysis. Multiplex PCR, Y Chromosome, STR 104 * Presenting Author B143 Development of a Haplotype Database for the AmpFlSTR® Yfiler™ PCR Amplification Kit Lisa M. Calandro, MPH*, Julio J. Mulero, PhD, Lori Hennessey, PhD, Lucas Rix, Charles Lawrence, Maryam Norouzi, and Dennis Reeder, PhD Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404 Attendees will receive an overview of the development of a haplotype population database for the AmpFlSTR® Yfiler™ PCR Ampliification Kit and the web-based tool for searching the haplotype database. This presentation will impact the forensic community and/or humanity by informing the forensic community of a new tool for the analysis of haplotypes obtained using the Yfiler™ PCR Amplification Kit. This presentation will discuss the development of a haplotype population database for the AmpFlSTR® Yfiler™ PCR Amplification kit. Attendees of the presentation will be provided with an overview of a webbased tool for searching the haplotype database. Statistical analyses of the population database and the detection of microvariants will be discussed. Analysis of sexual assault evidence using currently available autosomal amplification and typing systems is complicated by the inability to obtain male DNA profiles in the context of high quantities of female DNA. As a result, sexual assault evidence containing low numbers of spermatozoa or semen from azoospermic or vasectomized males may not be analyzed and, if analyzed, often does not yield male DNA profiles. Analysis of male-specific DNA markers can provide evidence profiles that may have probative or exculpatory value in such cases. Y STR analysis has proven to be a valuable tool for recovering information from evidence samples containing marginal amounts of male DNA. Due to the genetically linked structure of Y STR haplotypes, estimation of population frequencies for a particular haplotype requires analysis of population database samples using the complete set of markers used to analyze the forensic evidence sample. The AmpFlSTR Yfiler PCR Amplification kit may also be utilized for paternity analysis in situations involving male offspring, providing an additional tool for discrimination that will complement autosomal STR analysis. The AmpFlSTR Yfiler PCR Amplification kit features 17 male-specific DNA markers located on the Y chromosome. A small range of amplicon sizes, from 100-326 base pairs, has been achieved by labeling the amplified products with four fluorescent dyes, including 6-FAM, VIC, NED, and PET. The selected markers include the European minimal haplotype consisting of DYS19, DYS389I/II, DYS390, DYS391, DY6S392, DYS393 and DYS385a/b, the additional SWGDAM recommended loci, DYS438 and DYS439, as well as six additional loci, DYS437, DYS448, DYS456, DYS458, DYS635 (Y GATA C4), and Y GATA H4. The added loci significantly increase the discrimination potential of the Y haplotype. A haplotype database was developed in order to provide a useful tool for estimating Y haplotype frequencies in a variety of ethnic groups. Population samples from North American, European and Asian population groups were analyzed by various test site laboratories and compiled on the basis of ethnic group. The data are accessible via the Internet for haplotype searches. Features of the search tool include the ability to input evidence profiles both singly and in batches via automatic upload from Genotyper® software files. The tool is also compatible with data generated using GeneMapper® ID v3.2 software. A manual input mode allows entry of microvariant data resulting in the capability for all profiles to be searched and reported. Search outputs provide estimated frequencies for individual ethnic groups as well as for all population samples. Any haplotype matches to the forensic unknown will also be shown. As is the case for mitochondrial DNA, haplotype frequencies are estimated by counting the number of times the haplotype is seen in various populations. Therefore it is desirable to increase the size of the haplotype database where possible. Periodic updates to the database will reflect the ongoing analysis of additional population samples. Y STRs, Haplotype, Database * Presenting Author B144 The Amelogenin Sex Test: The Missing Y? Lim Eng Seng Simon, BS*, Health Sciences Authority, DNA Database Laboratory, 7 Maxwell Road, MND Building Annex B, #05-04, Singapore, 069111, Singapore; Syn Kiu-Choong Christopher, PhD, Health Sciences Authority, DNA Profiling Laboratory, 11 Outram Road, Singapore, 169078, Singapore; and Tan-Siew Wai Fun, MSME, Health Sciences Authority, DNA Database Laboratory, 7 Maxwell Road, MND Building Annex B, #05-04, Singapore, 069111, Singapore The goal of this presentation is to present the forensic community rare and interesting failures in the amelogenin sex test and to recommend alternative Y-locus (i.e., SRY) marker be routinely included in any sex test to avoid mistyping and for gender confirmation. In addition, more p-arm Y-STRs should be included in Y-STRs multiplex design to accommodate potentialy q-arm Y-chromosome deleted males. The amelogenin sex test is well established as the marker of choice for sex determination in forensic DNA typing work. This sex test is included in commercially available DNA typing kits. This presentation will impact the forensic community and/or humanity by highlighting two interesting cases that demonstrate the failure of the amelogenin sex test. Alternative Y-locus (i.e., SRY) marker should be routinely included in any sex test to avoid mistyping and for gender confirmation. In addition, more p-arm Y-STRs should be included in Y-STRs multiplex design to accommodate potentialy q-arm Y-chromosome deleted males. The oral presentation will highlight two interesting cases that demonstrate the failure of the amelogenin sex test: Case I: a male convicted offender was typed as “asexual” by AmpFlSTR® Identiflier™ DNA typing kit. Case II: a paternity test involving a family of 4 persons, where one of the 2 brothers showed a female genotype by AmpFlSTR® Profiler Plus™. In both cases, the cause of gender mis-identification using the commercial available amelogenin sex test found in the DNA typing kit was resolved using alternative amelogenin primers, sex determining region on the Y chromosome (SRY) primers, and Reliagene Y-Plex™ 12. In the first case, during routine DNA Database typing analysis, an amelogenin null male was encountered. Both amelogenin specific X and Y alleles was missing from his DNA profile, obtained using the AmpFlSTR® Identiflier™ DNA typing kit. Alternative amelogenin primers, which amplifies the X allele at 212 bp, and Y allele at 218 bp was used to verify the gender. Amplification using the alternative amelogenin primers, showed the presence of only the X allele. This indicates a point mutation at the primer binding position at the amelogenin X allele, which account for the failure in typing for the amelogenin X allele using the Identifiler™ DNA typing kit. However, the amelogenin Y allele still remain missing. The SRY gene amplification gave a positive result, which is the correct sex test result. Using the Reliagene Y-Plex™ 12 DNA typing kit, a complete Y-halpotype DNA profile was obtained. Results therefore, suggest that a deletion of the amelogenin gene must have occurred along the Y chromosome. In addition, a point mutation of the primer-binding site of the X allele used in the Identifiler™ DNA typing kit caused the total failure of the amelognin sex test, resulting in a null allele profile for the amelogenin locus. In the second case, a family of four, which includes the tested parents and their two sons, were genotyped using the AmpFlSTR® Profiler Plus™ DNA typing kit and their biological parentage relationship was established. Gender determination using the amelogenin sex test available in the DNA typing kit was correct for all, except for the older son, whose Y allele was missing from his DNA profile. Alternative amelogenin primers fail to amplify the Y specific allele as well. However, with SRY typing, the older brother was correct for his gender. Reliagene Y-Plex™ 12 was then used to determine the Y-STRs profiles of the biological father and his two sons. The Y-STRs haplotype of the biological father and his younger son was found to be both complete and consistent with one another. For the older 105 son, only the Y-chromosomal STR marker DYS393 and the X allele of the amelogenin locus were typed. The DYS393 allele was found to be consistent with that of his father and younger brother. The results, therefore indicate that a deletion polymorphism possibly, spanning a major part of the Y-chromosome from Yp11.31 on the short arm up to Yq11.221 on the long arm has occurred. This deletion event, would explain his missing Y allele from the amelogenin maker using the Profiler Plus™ DNA typing kit. In addition, the inclusion of more Y-STRs on the p-arm of the Y chromosome for e.g. DYS453, DYS446 and DYS456 in Y-STRs multiplex design should be considered. In this way, Y chromosome with deleted qarm can still allow discriminating Y-STRs information to be obtained. Amelogenin, Gender Identification, Y-STRs B145 Genotyping of Nuclear Loci From Telogenic Hair Shafts Using mini-STRs Michael D. Coble, PhD*, Peter M. Vallone, PhD, and John M. Butler, PhD, National Institute of Standards and Technology, 100 Bureau Drive, Mail Stop 8311, Gaithersburg, MD 20899-8311 After attending this presentation, attendees will learn an approach to increasing the successful typing of nuclear DNA from telogenic hair samples using miniSTRs for both CODIS and non-CODIS markers will be described. This presentation will impact the forensic community and/or humanity by demonstrating how the successful typing of even a small number of nuclear loci from shed hairs can greatly increase the forensic discrimination of the sample compared to mtDNA testing alone, where a significant number of common types are present in the population. Forensic DNA analysts often perform short tandem repeat (STR) typing on highly degraded biological material and then turn to mitochondrial DNA (mtDNA) testing, which is less variable but more likely to obtain a result due to higher copy numbers in cells, if many or all of the STRs fail. MtDNA typing of hair shafts is a particularly important application as shed hairs are commonly found as sources of evidence. Currently, forensic hair comparisons of evidentiary and reference specimens are based upon a set of morphological characteristics. These analyses tend to be subjective, relying on the experience and judgment of the examiner.1 A number of studies have demonstrated that successful analysis of degraded DNA specimens from mass disasters or forensic evidence improves with smaller sized polymerase chain reaction (PCR) products.2 By moving PCR primers closer to the STR repeat region, it is possible to obtain fully concordant results to the commercial kits while improving successful analysis of degraded DNA with smaller PCR products or miniSTRs2. However, many of the CODIS core loci have large allele ranges (e.g., D21S11 and FGA) that make it impossible to create small PCR products. The authors are also going beyond the CODIS core loci and examining a battery of new potential STR loci that can be made less than 100 bp in size and would therefore be helpful in testing highly degraded DNA samples.3 Methods and Materials: Hairs were digested using either a standard micro-tissue grinding protocol or a complete digestion protocol.4 DNA template was purified with phenol/chloroform/water followed by microcon concentration or via binding/elution using a Qiagen column. PCR reactions using STR miniplex markers of CODIS markers (TH01, FGA, D18S51, D16S539) along with the D2S1338 marker and Amelogenin were evaluated. An additional miniplex containing non-CODIS markers (D10S1248, D14S1434, and D22S1045) was also evaluated. Summary of Results: A set of two miniSTR multiplexes containing CODIS and non-CODIS markers have been evaluated for their ability to genotype degraded DNA. A number of the miniSTR markers were used to successfully type nuclear DNA from hairs belonging to multiple individuals. 106 Conclusions: The selection of STR loci that have a narrow allele range (e.g., less than 50 bp) and can be made smaller than 100 bp works well with degraded DNA samples. The successful typing of even a small number of nuclear loci from shed hairs can greatly increase the forensic discrimination of the sample compared to mtDNA testing alone, where a significant number of common types are present in the population. References: (1) Bisbing, R.E. (1982). The forensic identification and association of human hair. in Forensic Science Handbook, R. Saferstein, ed. (New Jersey: Prentice Hall Regents), pp. 184-221. (2) Butler, J.M., Shen, Y., McCord, B.R. (2003) The development of reduced size STR amplicons as tools for analysis of degraded DNA. J. Forensic Sci., 48(5): 1054-1064. (3) Coble, M.D. and Butler, J.M. (2004) Characterization of new miniSTR loci to aid analysis of degraded DNA., submitted. (4) Hellmann, A., Rohleder, U., Schmitter, H., Wittig, M. (2001) STR typing of human telogen hairs – a new approach. Int. J. Legal Med., 114: 269-273. Short Tandem Repeat DNA Typing, Degraded DNA, Reduced Size PCR Products B146 Optimized Extraction of Nuclear DNA From Hair Shafts: Part 2 Sheila M. Estacio, MS*, Stacey Murnyak, BS*, Zoran Budimlija, MD, PhD, and Mechthild Prinz, PhD, Office of Chief Medical Examiner, Department of Forensic Biology, 520 First Avenue, New York, NY 10016 After attending this presentation, attendees will learn a method to extract nuclear DNA from hair shafts (without roots) on both head and pubic hairs Traditionally, it is known that nuclear DNA from hair shafts is minimal to non-existent in the absence of a root or adhering epithelial cells. This method proves that nuclear DNA can be obtained from hair shafts. This method can be implemented in forensic DNA labs equipped mainly for STR DNA testing. It can also be used in labs also online for forensic mtDNA testing to use in conjunction with the STR profile to increase the statistics on the profile obtained whether it be for use in criminal cases or for missing persons. The profile or profiles can then be uploaded to CODIS. This research initially began as work to test hair samples recovered from the World Trade Center Disaster and the hair samples received as references provided by the families as well as direct reference hair samples from the victims. The experiments during Part 1 explored decontamination methods, application of various enzymes for digestion, enzyme concentrations, different volumes of incubation buffer, amplification strategies, and varied run parameters on the ABI Prism 3100 Genetic Analyzer. Part 2 focuses on comparing two modified extraction protocols using Promega’s DNA IQ versus a direct lysis extraction. Head and pubic hairs were collected. The hairs were cut to ensure that no roots were present. If the hairs were not cut, they were examined macroscopically and microscopically to ensure that no roots were present. The hairs were washed using 5% Terg-a-zyme, 0.9% NaCl solution, and 100% ethyl alcohol and air dried overnight. The dried head hair was milled into a powder using the Spex Certi-Prep Freezer Mill and measured out in 20mg amounts. The pubic hairs were washed in the same way and each cut to 2.5 cm in length. The pubic hairs were pooled together to 12.5 cm and 25 cm lengths. The hairs were then incubated in 1.8 mg/ml Proteinase K, 0.25M DTT, 0.5% SDS, and DNA IQ incubation buffer containing 10mM Tris (pH 8.0), 100mM NaCl, and 50mM EDTA overnight at 56°C. The volume of incubation buffer varied depending on the type of hair sample-milled hair or pubic hair strands. For the milled head hair, the supernatant was taken and divided evenly between the optimized DNA IQ and direct lysis * Presenting Author extraction. For DNA IQ, the recommended amount of Resin was used followed by a Microcon 100 cleanup. For the direct lysis extraction, the extract was purified through either a Bio-Rad Micro Bio-Spin Chromatography or a Centri-Sep column followed by a Microcon 100 clean-up. The amount of DNA was quantified using a real-time PCR Alu-based assay (Nicklas and Buel). Attempts at quantifying the amount of nuclear DNA were initially done using the slot-blot method (Quantiblot). As expected, no DNA was detected since the sensitivity of Quantiblot is insufficient to less than 150 picograms. The use of the real-time technology is advantageous for quantifying the low level amount of nuclear DNA in the hair shaft extraction, to detect if melanin is inhibiting amplification, to check the “success” of the positive and negative controls, and also to predict the success of STR amplification. Results from head hair and pubic hair are compared as well as the differences in the amount of DNA recovered from blond, brown, and black colored hairs. Initial results for 20 mg of milled head hair show that naturally blond hair yields a stronger signal in real-time PCR than brown or black colored hairs for both extraction methods. This further proves that melanin can act as a PCR inhibitor. Comparing the two extraction methods, the optimized DNA IQ protocol is generally more successful at “capturing” about three times more the amount of DNA than by using the direct lysis method. Initial STR data based on Promega’s PowerPlex® 16 show the expected amount of DNA degradation with drop out of the larger loci. Future work will discuss the results of the testing on pubic hair strands, additional STR results using PowerPlex®16, and the viability of the extract and success for mtDNA sequencing. Hair Shafts, Extraction, Nuclear DNA B147 Evaluation and Quantification of Nuclear DNA From Human Telogen Hair Kerry L. Opel, MA*, and Bruce R. McCord, PhD, Florida International University, University Park Department of Chemistry and Biochemistry, 11200 Southwest 8th Street, Miami, FL 33199; and Erica L. Fleishaker*, and Weston Garber, Ohio University, Department of Chemistry and Biochemistry, 136 Clippinger Laboratories, Athens, OH 45701 After attending this presentation, attendees will be familiar with the procedures utilized in the extraction, quantification and amplification of nuclear DNA from single telogen hairs. This presentation will impact the forensic community and/or humanity by providing information on the practicality of extracting and amplifying nuclear DNA from human telogen hair. This paper presents data on the extraction and quantification of nuclear DNA from naturally shed hairs such as those that would be present at crime scenes. Additional goals of this project are: to determine the significance of exogenous DNA on hair; to evaluate method for the removal of exogenous DNA from the hair; and to evaluate the state of degradation of the DNA using real time PCR. The quantified DNA was amplified using small amplicon STR kits (Miniplexes) and a commercial STR typing kit. Many experiments have been published on the extraction and amplification of DNA from hair. While some success has been reported for amplification of DNA from hair, accurate quantification of the extracted DNA has been a problem. Before the use of real time PCR for quantification, DNA analysts have relied on methods which lack the sensitivity required to detect the minute amounts of DNA found in hair. Therefore, information on the actual amount of nuclear DNA that can be recovered from hair is scarce. The use of real time PCR for quantification allows for detection of DNA in the picogram range, and this method is suitable for a study of the amount of DNA that can be recovered from telogen hairs. In order to ensure that the DNA quantified comes from within the hair, methods that can remove exogenous DNA must be utilized. The use of differential extraction is a method that allows removal of epithelial cells * Presenting Author from the surface of hair, and also allows for the evaluation of the impact of exogenous DNA through quantification of the recovered DNA and evaluation of contamination by amplification of the exogenous DNA. Direct evaluation of the state of degradation of DNA can normally be accomplished through agarose gel separation and ethidium bromide staining. However, this method required microgram amounts of DNA. When only picogram or nanogram amounts are recovered, real time PCR can be used to determine the number of small and large fragments through amplification with different primers. Comparison of the amounts of large and small fragments will indicate if degradation is present in the sample. When DNA recovered from hair is degraded, success of amplification of the DNA using multiplex kits with large amplicons can be limited. Instead, Miniplex kits specifically designed for use with degraded DNA can be used as an alternative for such samples. In this project, DNA was extracted from telogen hairs (3 cm in length) from numerous volunteers. Eight to nine hairs from each individual were extracted using a published protocol consisting of a calcium based extraction buffer system with DTT, phenol chloroform separation, and Microcon® YM-30 filtration. The extracted DNA was quantified by real time PCR using the Corbett Rotor-Gene 3000, a 124 base pair Alu amplicon, and SYBR Green I dye. In order to evaluate the amount of exogenous DNA contamination, a differential extraction buffer without DTT was used to remove epithelial cell DNA, and the DNA removed was extracted, quantified, and amplified with the Miniplex kits. Samples with the highest concentrations were concentrated further using Microcon® filters. The concentrated extracts were quantified with two different sized Alu primers (124 and 280 base pairs) and the quantification results were compared to determine the state of degradation of the extracted DNA. The concentrated extracts were then amplified with the Miniplex kits, and the results of the quantification were compared to the profiles to determine the effect of the degradation on genotyping. The amount of DNA recovered from hair using this method varies greatly, with a range of 1-90 pg/ L. The amount of exogenous DNA present in hair also varies, and the exogenous DNA appears to have little contamination from outside sources. DNA recovered from hair does show degradation upon evaluation by real time PCR, but the DNA can be successfully amplified using reduced size STRs. Nuclear DNA, Telogen Hair, Real Time PCR Quantification B148 Applications of Visible and Near Infrared Chemical Imaging to the Analysis of Forensic Evidence Gemma Payne, BSc, and Brian Reedy, PhD, University of Technology, Sydney, Centre for Forensic Science, PO Box 123, Broadway, NSW 2007, Australia; Chris Lennard, PhD*, Australian Federal Police, Forensic Services, GPO Box 401, Canberra, 2601, Australia; David Exline, MSFS, R.J. Lee Group, Inc., 350 Hochberg Road, Monroeville, PA 15146; Rebecca Schuler, BSc, ChemImage Corporation, 7301 Penn Avenue, Pittsburgh, PA 15208; and Claude Roux, PhD, University of Technology, Sydney, Centre for Forensic Science, PO Box 123, Broadway, NSW 2007, Australia Attendees will gain an understanding of chemical imaging and the general application of this new technology in the field of forensic science. With technology continuously evolving, it is important to explore new developments that may prove to be superior to techniques currently in use in forensic laboratories. This presentation will impact the forensic community and/or humanity by demonstrating how chemical imaging offers a higher discriminating power and greater sensitivity compared to conventional digital imaging. This oral presentation will describe the theory of chemical imaging and demonstrate how this emerging technology has enormous potential in the forensic analysis of materials when compared to traditional digital imaging techniques. 107 Chemical imaging combines molecular spectroscopy and digital imaging, providing both spatial and spectral information of materials. Light intensity is detected as a function of wavelength as in spectroscopy, and is also detected as a function of location as in conventional imaging to form a data set. The three-dimensional data set has a fully resolved image at each individual wavelength and a full spectrum at each individual pixel. Valuable information can then be extracted from the large data sets through the utilization of specialized software designed for chemometric and image analysis. Common processing tools include zero offset, normalization and principal components analysis. This promising technology has a wide variety of applications in many scientific and industrial fields, one of which is in forensic science. Chemical imaging offers many advantages for forensic science applications. Over a short analysis time, information on morphology, composition, structure and concentration can be obtained and displayed side-byside. It is also a non-destructive technique that utilizes well-accepted microscopy and spectroscopy methods, with little or no sample preparation required. The liquid crystal tunable filter (LCTF), the basis of the chemical imaging instruments considered here, is extremely advantageous as it replaces the often numerous detection filters needed. The LCTF is able to perform analysis with less than 1 nm increments, which greatly increases the sensitivity capabilities of the analysis. The VIS/NIR ChemImage ‘Condor’ Macroscopic Chemical Imaging system, as used in this research, consists of an electro-optical tunable filter system with a charged couple device (CCD) camera on a macroscopic platform. The total spectral range of the VIS/NIR Condor is from 400 nm to 1100 nm. This research project is focused on establishing the potential for the use of the VIS/NIR Condor for forensic evidence analysis with special focus on the analysis of inks on questioned documents. Over 110 pens have been sampled (specifically blue and black ballpoint and roller-ball pens). Chemical imaging analysis using the VIS/NIR Condor was conducted on inks deposited on white paper. Resulting data sets were then processed using the software to distinguish between the different inks. Traditional optical techniques currently in use for ink analysis were used to obtain comparison data. Other avenues of research include firearm propellant analysis, latent fingerprint detection, and textile fiber comparison. Unfired and fired firearm propellant was examined using the VIS/NIR Condor with the objective of determining if fluorescence can be used for the detection and identification of propellant grains. Latent fingerprints on porous and nonporous surfaces were examined before and after common reagent treatment to establish if additional ridge detail could be detected using the chemical imaging system. Red and black textile fibers were examined in transmittance, reflectance and fluorescence mode to ascertain if Chemical Imaging produced superior discriminating powers compared to traditional techniques. All evidence types examined produced extremely promising results. The chemical imaging technology was able to distinguish a greater number of inks and fibers. Propellant grains on a target could be traced to the brand that was used in the shooting. The VIS/NIR Condor proved to be an advantage when examining latent fingerprints, especially weak marks or marks on surfaces that produce highly luminescent backgrounds. Untreated latent fingermark ridge detail could also be detected in some cases. This is an ongoing research project that will be expanded to include a range of evidence categories to fully validate the technology. Initial results show that chemical imaging has great potential in the analysis of forensic evidence including questioned documents, firearm propellants, fingerprint detection and textile fibers. Chemical Imaging, Ink Analysis, Trace Evidence 108 B149 Combinatorial Optimization of the Extraction of Dyes From Textile Fibers Amy R. Stefan, BS*, Christopher R. Dockery, BS, Alexander A. Nieuwland, PhD, Samantha N. Roberson, James E. Hendrix, PhD, and Stephen L. Morgan, PhD, University of South Carolina, Department of Chemistry and Biochemistry, 631 Sumter Street, Columbia, SC 29208; and Mark L. Miller, PhD, FBI Laboratory, Counterterrorism and Forensic Science Unit, FBI Academy, Quantico, VA 22135 The goal of this presentation is to report the development of automated procedures for the extraction of dyes from textile fibers and analysis of dye extracts by spectroscopic methods for possible adaptation to forensic casework. This presentation will impact the forensic community and/or humanity by demonstrating how optimization of extraction conditions can provide the analyst with the maximum amount of dye, possibly providing additional discrimination between forensic fiber evidence. This presentation will report the use of a Beckman-Coulter BioMek® 2000 laboratory automation workstation to systematically investigate a combinatorial approach to the development of micro-scale extraction techniques for a broad range of dye classes from a range of textile fiber types. The laboratory workstation was programmed to extract dyes from small fiber samples. The resulting extracts were used to quantitatively determine the extraction efficiencies of different extraction solvents. Details of the programming and extraction technology will be presented along with experimental results, conclusions, and potential impact on forensic science. Use of an automated workstation to extract dyes from fibers presents several advantages. Both speed of analyses and sample throughput are drastically increased versus manual extraction. In addition, the automated workstation gives the ability to program combinatorial experiments for the determination of the solvent mixtures that will provide the most efficient extraction conditions possible for a given fiber-dye interaction. Extraction efficiencies were quantitatively determined via designed combinatorial experiments and generation of surface response models. Knowledge of the chemistry of fibers and fiber dyes is important to developing extraction techniques. Depending on the chemistries of the dye-fiber interaction, dyes may be loosely associated with fibers (e.g., direct dyes), bound by salt linkages (e.g., acid dyes), covalently bound to the fiber (e.g., reactive dyes), or simply dispersed as finely divided pigments in the fiber (e.g., disperse dyes). Dyes may also be applied during melt spinning of thermoplastic fibers (e.g., pigment coloration of nylon, polyolefins and polyester) or adhered to the fiber surface with adhesives (e.g., pigment dyeing of bedding and apparel fabrics). The following fiber/dye combinations have been successfully extracted using automated extraction techniques: acid dyes on nylon, basic dyes on acrylic, direct, vat, or reactive dyes on cotton, and disperse dyes on polyester. Acid dyes usually contain sulfonic or carboxylic acid groups that form salt linkages to the nylon. Basic dyes on acrylics, also called cationic dyes, contain basic groups that form salt linkages to the acrylic. Cotton is usually dyed using direct, vat, or reactive dyes. Direct dyes form hydrogen bonds with the cellulose, and usually contain sulfonic or carboxylic acid groups, rendering them soluble in water. Vat dyes, which are usually quinone structures, are reduced to their water-soluble leuco form for application to the fibers, and re-oxidized to form insoluble pigments to improve wash fastness. Reactive dyes contain functional groups that form covalent bonds to the cellulosic hydroxyl groups. Lastly, disperse dyes on polyester are finely divided organic pigments dispersed throughout the fiber. Dyes were extracted from fibers using the automated workstation, and loaded into a 96-well plate system. In this 96-well plate system, each well contains a 500- L glass insert. A Teflon liner was placed between the glass inserts and a plastic lid to minimize evaporation of the extraction solvent during high-temperature extractions. To further minimize evaporation of solvents during extraction, the assembled plate was clamped in an alu- * Presenting Author minum press to improve the seal between the Teflon liner and the glass inserts. Extracted dyes were analyzed by capillary electrophoresis (CE) with a PACE-MDQ CE system using diode array detection. Optimization of extraction conditions provides the analyst with the maximum amount of dye, possibly providing additional discrimination between forensic fiber evidence. extraction and further analysis may only be considered if additional discrimination is needed after forensic microspectroscopy and microspectrophotometry has been conducted. Discrimination of Fiber Dyes, Capillary Electrophoresis, Mass Spectrometry Textile Fibers, Dyes, Automated Extraction B150 Capillary Electrophoresis/Diode Array Detection/Mass Spectrometry for the Forensic Analysis of Fiber Dyes B151 UV-Visible and Fluorescence Microspectrophotometry for the Forensic Analysis of Fluorescent Brighteners on Textile Fibers Alexander A. Nieuwland, PhD*, Brandi L. Clelland, BS, James E. Hendrix, PhD, and Stephen L. Morgan, PhD, University of South Carolina, Department of Chemistry and Biochemistry, 631 Sumter Street, Columbia, SC 29208; and Mark L. Miller, PhD, FBI Laboratory, Counterterrorism and Forensic Science Unit, FBI Academy, Quantico, VA 22135 Stephen L. Morgan, PhD*, Alexander A. Nieuwland, PhD, Elizabeth M. Enlow, BS, and James E. Hendrix, PhD, University of South Carolina, Department of Chemistry And Biochemistry, 631 Sumter Street, Columbia, SC 29208; and Edward G. Bartick, PhD, FBI Laboratory, Coutnerterrorism and Forensic Science Research Unit, FBI Academy, Quantico, VA 22135 The goal of this presentation is to determine the usefulness of capillary electrophoresis-diode array detection-mass spectrometry (CE-DAD-MS) on extracted fiber dyes for forensic fiber discrimination. This presentation will impact the forensic community and/or humanity by demonstrating CE-DAD-MS of extracted fiber dyes represents a supplement to microscopy and microspectrophotometry for the forensic discrimination of fibers. Fiber evidence is frequently used in forensic science to associate a suspect to a victim or crime scene. At times “questioned” fibers are collected from the crime scene and “known” fibers are collected from suspects for comparison. Evidence fibers are collected through a combination of picking, scraping, vacuuming, and sometimes taping clothing and areas of the crime scene. They are often mounted in a mounting material on microscope slides for comparison and storage. Once the fibers have been collected, questioned and known fibers are compared using a series of microscopic techniques to determine whether or not the fibers could have come from the same source. The first of these methods is often polarized light microscopy (PLM). Using PLM, the generic fiber type (polyester, acrylic, nylon, cotton, etc.) is determined, and color, fiber cross-sectional shape and fiber thickness are compared. This analysis is often followed by fluorescence microscopy and UV-Vis and fluorescence microspectrophotometry. If spectra of the known and questioned fibers are consistent, the hypothesis that the fibers originate from a common source should not be rejected. Additional discrimination may be achieved by extraction of the fibers dyes followed by chromatographic analysis. CE-DAD-MS was chosen as the analysis technique following extraction because of its high sensitivity and selectivity. Fiber dyes were extracted using a Beckman-Coulter Biomek 2000 liquid sample-handling robot. Extracted dyes were analyzed using a Beckman-Coulter P/ACE MDQ capillary electrophoresis system coupled to a Micromass Q-TOF micro mass spectrometer. An external fiber optic light source with a xenon lamp was used as the light source for the diode array detector to enable simultaneous DAD and MS detection. This setup provides an improved signal to noise ratio in the visible region and a better agreement between peaks in the DAD and MS electropherograms. CE-DAD of small molecules has often employed sodium acetate or phosphate buffers with cationic surfactants or cyclodextrins as buffer additives. However, because of the requirements of the electrospray ionization process, non-volatile buffers and buffer additives should be avoided in CEMS. CE-DAD-MS methods for the analysis of cationic dyes from acrylics, direct, vat, and reactive dyes from cotton, acid dyes from nylons, and disperse dyes from polyester were developed. Overall, CE-DAD-MS of extracted fiber dyes represents an alternative approach for the forensic discrimination of fibers. However, because of the comparatively high cost and time requirements of CE/MS, The goal of this presentation is to determine the usefulness of UV-Vis and fluorescence microspectrophotometry of fluorescent brighteners on fibers for forensic fiber discrimination. This presentation will impact the forensic community and/or humanity by demonstrating how fluorescent brighteners can be important for discriminating among seemingly similar white fibers. The absorbance and fluorescence of fluorescent brighteners on acrylic, cotton, nylon and polyester fibers can be detected and differentiated in Permount without significant interference from absorbance or fluorescence from the mounting material. C Different fluorescent brighteners from the same chemical subclass (e.g., pyrazolines) showed similar spectra that could also be differentiated. Fiber evidence is frequently used in forensic science to associate a suspect to a victim or crime scene. “Questioned” fibers are collected from the crime scene and “known” fibers are collected from the suspect. Evidence fibers are collected through a combination of picking, scraping, vacuuming, and sometimes taping clothing and areas of the crime scene and are mounted in a mounting material on microscope slides for comparison and storage. Once the fibers have been collected, questioned and known fibers are compared using a series of microscopic techniques to determine whether or not the fibers could have come from the same source. The first of these methods is often polarized light microscopy (PLM). Using PLM, the generic fiber type (polyester, acrylic, nylon, cotton, etc.) is determined, and color, fiber cross-sectional shape and fiber thickness are compared. This analysis is often followed by fluorescence microscopy, and UV-Vis and fluorescence microspectrophotometry. If spectra of the known and questioned fibers match, the hypothesis that the fibers originate from a common source should not be rejected. Many fibers not only contain dyes that absorb visible light (400-700 nm), but also optical brighteners, which absorb ultra-violet light (around 360 nm) and re-emit light as fluorescence within the visible spectrum (around 440 nm). These fluorescent dyes mask “yellowness” and can make fabrics appear “whiter than white.” Most of the attention in the analytical literature has been focused on fluorescent brighteners from detergents, with little attention being paid to fluorescent brighteners applied by the textile dyer. Fluorescent brighteners are often the only dyes present on white fibers. They are added to the melt during manufacture of staple polyester for blending with cotton. Textile dyers add fluorescent brighteners to impart whiteness to woven and knit products. Everyone who launders his/her clothing also adds fluorescent brighteners because they are in almost every brand of laundry detergent. Fluorescent brighteners can therefore be important for discriminating among seemingly similar fibers. For this paper, nine different fluorescent brighteners from six different chemical subclasses were compared: one coumarin, two distyrylbiphenyls, one heterocycle, two pyrazolines, two * Presenting Author 109 stilbenes, and one thiophene oxazole. One objective of this study was to determine whether Permount®, a popular mounting medium for UV/vis spectrophotometric characterization, would result in loss of discriminatory spectral data for fibers dyed with fluorescent brighteners. To answer this question, slides of polyester, acrylic and nylon 6 fibers, with and without fluorescent brightener, were prepared using Permount and glycerin as mounting media on glass and quartz. Additionally, the possibility of discriminating between seemingly similar white fibers with different fluorescent brighteners based on their UV-Vis and fluorescence spectra was assessed. The absorbance and fluorescence of fluorescent brighteners on acrylic, cotton, nylon and polyester fibers can be detected in Permount without significant interference from absorbance or fluorescence from the mounting material. Comparison of fluorescence spectra by visual analysis and multivariate data analysis showed that fluorescence microspectroscopy could differentiate between all 9 different fluorescent brighteners used in this study. Different fluorescent brighteners from the same chemical subclass (e.g., pyrazolines) showed similar spectra that could nevertheless still be differentiated. In the case of cotton and nylon, fluorescent brighteners from detergents may increase the discrimination. Fluorescent brighteners contained in the detergent tested, however, did not show substantivity for polyester and acrylic fibers. Overall, UV-Vis and UV-fluorescence microspectroscopy using Permount is a viable approach for forensic discrimination of white fibers. For best results, forensic investigators should use as thin a layer of Permount as possible, and routinely take the 365 nm excitation fluorescence spectra of white fibers that show an absorbance peak around 360 nm. Further discrimination may be achieved by extraction of fluorescent brighteners from fibers dyes followed by chromatographic analysis. chemically react with the dyed fiber, be relatively easy to use, adhere and harden rapidly for permanent storage (if desired), have high optical clarity with no formation of bubbles or crystals, have a refractive index near 1.5 for performing polarized light microscopy (PLM), be soluble in a non-toxic solvent to facilitate recovery of the fiber from the medium, be colorless, be non-fluorescent, not yellow or shrink with age, and be inexpensive. While no mountant possesses all of these ideal properties, a suitable mountant must have most of these characteristics, and of particular importance is that it must not react with the textile fiber. The authors have evaluated three different mounting media (glycerin, Norland Optical Adhesive 65, and Permount®) for the forensic discrimination of fibers by UV-VIS and UV/fluorescence. Subject to a few working recommendations, it has been found that Permount to be a suitable mountant for analyses. A physical database containing over 500 dyed and undyed textile fibers has been developed with contributions from textile companies in the southeastern United States. All fibers have been characterized by polarized light microscopy, UV/Visible, and fluorescence microspectroscopy. A total of 25,000 UV-vis and fluorescence spectra have been acquired. Visual comparisons among the fiber spectra from the database have been supplemented by multivariate statistical analysis to confirm the statistical validity of discrimination observed. Overall, UV/Vis and fluorescence microspectrophotometry are valuable tools for the discrimination of fibers, in particular for discrimination of fibers of similar color but different dye composition. The fibers and associated spectra in the database are a useful tool for fiber comparisons in casework and in quality control and training of analysts. Recommendations will be made for approaches which produce the most discriminating data to ensure that the limited time and resources available in forensic laboratories are most efficiently applied. Fibers, Fluorescent Brighteners, UV-Vis/Fluorescence Microspectrophotometry Fibers, UV-Visible Microspectrophotometry, Fluorescence B152 Forensic Discrimination of Dyed Textile Fibers Using UV-Vis and Fluorescence Microspectrophotometry Stephen L. Morgan, PhD, Alexander A. Nieuwland, PhD, Christopher R. Mubarak, PhD, James E. Hendrix, PhD, Elizabeth M. Enlow, BS, and Bryan J. Vasser, University of South Carolina, Department of Chemistry and Biochemistry, 631 Sumter Street, Columbia, SC 29208; and Edward G. Bartick, PhD*, FBI Laboratory, Counterterrorism and Forensic Science Research Unit, FBI Academy, Quantico, VA 22135 The goal of this presentation is to evaluate the relative discriminating power of visible, UV/visible, and UV/fluorescence spectrophotometry for the characterization of dyed textile fibers. This presentation will impact the forensic community and/or humanity by demonstrating how UV/Vis and fluorescence microspectrophotometry are valuable tools for the discrimination of fibers, in particular for discrimination of fibers of similar color but different dye composition. The fibers and associated spectra in the database are a useful tool for fiber comparisons in casework and in quality control and training of analysts. Fibers are ubiquitously found as trace evidence in crimes of personal contact, such as homicide, assault, sexual offenses, and hit-and-run accidents. Ultraviolet-visible (UV-VIS), and fluorescence microspectrophotometry (MSP) offers direct, relatively inexpensive, and informative means of characterizing dyed textile fibers. Research in the use of UV-VIS and fluorescence MSP has multiple objectives: improving the forensic discrimination of fibers by defining protocols for the most discriminating approaches, validating data analysis methods, and providing a tested database of spectra from dyed and undyed textile fibers. Fiber samples for forensic microspectrophotometry are typically mounted in a mounting medium (or mountant). The mountant must not 110 B153 A Study of Fabric Frequency Robin T. Stoehr, BS*, and Max M. Houck, MA, West Virginia University, Forensic Science Initiative, 886 Chestnut Ridge Road, PO Box 6216, Morgantown, WV 26505-6216 Attendees will learn an example of fiber content and frequency distribution among college students over time. The majority of reported victims and offenders of sexual assault are college aged, with the rape victimization highest among 16 – 19-year-olds. In 2002, nearly 4,000 sexual assaults were reported on college campuses nationwide. These crimes involve considerable contact between the perpetrator and the victim, allowing fibers to provide information about the crime scene as well as the vehicle used to transport a victim. This presentation will impact the forensic community and/or humanity by demonstrating data from the current project that helps to answer questions such as which fibers are more prevalent than others? Which fibers are rare? Are these fibers rare, or just not as common within this particular population? Population studies provide forensic scientists with a basis for understanding and interpreting the evidence they analyze, especially as it varies across geography, culture, and economic status. This paper will present the work done at West Virginia University to determine the relative frequencies of fiber types and content in a collegeaged cohort over time. The research was conducted throughout the month of April 2004. On a voluntary basis, 60 students were asked to describe their clothing three days a week. A variety of garment types (i.e., shirt, sweater, t-shirt, jeans, pants, etc.) were available to choose from when examining up to two top garments and one bottom garment. This excluded outer garments, such as jackets, coats, or slickers. Using the Kelly system, primary, secondary, and tertiary colors of each garment in addition to the modifiers light and dark were recorded. The students were asked to record the fiber type and respective percentage of each garment as reflected on the garment label(s). * Presenting Author This data was compiled to observe the totality of each garment type, primary, secondary, and tertiary colors, and fiber type and percentage. Out of the 619 #1 upper garments, 347 were t-shirts; 541 or 87.4% stated cotton as the primary fiber type. Polyester and acrylic were the two other major fiber types with 6.0% and 2.4%, respectively. Color distribution was more even with the most frequent being white throughout primary, secondary, and tertiary colors. The #2 upper garments were also primarily t-shirts at 126 out of the 244 total shirts observed. The fabric distribution was nearly the same as upper garment #1, with cotton being the most prevalent followed by polyester and acrylic including a high frequency of white shirts. The bottom garments showed 398 out of the 620 were blue jeans, or denim. Primary fiber types show that cotton was 93.3% of the total fiber type among bottom garments. Polyester comes in next with 4.2% of the total primary fiber type. Blue was the most prevalent primary color of all bottom garments. White cotton is the primary fabric type among the sampled group of college students; white cotton is often noted for its lack of specificity in fiber examinations. Excluding white cotton, polyester and acrylic in a variety of colors are the most prevalent. The frequency of fibers such as acetate, linen, and lyocel are non-existent in upper garments. Linen does not seem to be popular among the college students sampled. Fashion plays a large role in the fiber and fabric choices of this demographic and this influences the potential evidence available to forensic scientists whose casework involves college students in a temperate climate. Fiber Analysis, Fabric Content, Fiber Distribution B154 The Instrumental Analysis of the Volatile Organic Compounds Present in Human Scent by SPME-GC/MS and the Evaluation of Scent Preservation Methods Allison M. Curran, BS*, José R. Almirall, PhD, Adee A. Schoon, PhD, and Kenneth G. Furton, PhD, International Forensic Research Institute, Florida International University, University Park, Miami, FL 33199 Attendee will learn about the commonalities and the differences in human scent profiles across different individuals. This presentation will impact the forensic community and/or humanity by demonstrating the variability of human scent profiles and allowing for the better utilization of human scent evidence as an investigative tool in criminal cases. Human scent evidence and scent identification canines have become more commonly used by the law enforcement community for investigative purposes in recent years. Canines, Canis lupus var. familiaris, have demonstrated the ability to discriminate between individuals based on their odor. For these identifications to be valid, the hypothesis about human scent is that each person has a unique odor that is stable over time. Scientific research into the ability of canines to distinguish between individuals based on their scent supports this theory. The research presented here also supports the hypothesis of the uniqueness of human scent. Solid Phase Microextraction Gas Chromatography Mass Spectrometry (SPME-GC/MS) has proven to be a viable method for the extraction, separation, and identification of the volatile compounds which comprise human scent and various types of compounds have been identified, such as organic fatty acids, alcohols, aldehydes, and ketones * Presenting Author The majority of the scientific research into human odor has been conducted on sweat collected from the axillary (armpit) area and the feet by the cosmetics industry. The production of human odor is a complicated process which has yet to be fully understood. A number of factors make the axillary region a good odor producing area in the human body: (1) the contents of the apocrine gland secretions may serve as bacterial substrates; (2) moisture is available from the eccrine glands; (3) there is a resident population of bacteria to transform non-odorous substances to odorous substances; and (4) the presence of axillary hair may aid in the dispersion of the odor. The human body has 2-4 million sweat glands distributed over its surface. Sweating is the process of releasing a fluid secretion on the body’s surface in an effort to control temperature. The human axillary (armpit) region is the area of the body where the largest collection of sweat glands in both size and number are located. Apocrine, eccrine, and sebaceous glands, which are the major glands responsible for the secretion of “sweat,” are all present in the axillary region of the body. Forensically, odor collected from the hand is also of great interest. The use of detector dogs for “human scent lineups” has been utilized in European countries, including the Netherlands, Poland, Russia, Belgium, Germany, Denmark, and Hungary for decades. A “human scent lineup” is an identification based on a canine matching the human scent collected from a crime scene to a possible suspect. The process for conducting a “human scent line-up” in the Netherlands begins when scent evidence is collected at a crime scene, packaged, and preserved. When a suspect is taken into custody he or she may be asked to submit to a “human scent lineup.” The suspect then holds a metal bar in his hands for a period of time, and this metal bar is collected. This metal bar from the suspect along with metal bars that have been held by other individuals and collected at random throughout the population are set up in a sterile room, where the law enforcement certified canine is then exposed to the scent evidence, and allowed to work the line-up of hand scented metal bars independently. Scent identification indicates an association between the suspect and the scent evidence. Canines performing “human scent line-ups” are utilizing hand odor for their distinctions between individuals. This paper discusses the use of SPME-GC/MS for the analysis of scent samples collected from both the hand and armpit region of individuals differing in age, sex, and race. The stability and reproducibility of an individual scent profile over time for the purpose of creating a baseline for human odor will also be discussed. The common compounds determined between individuals and their relative ratios across individuals will be presented along with some compounds that are uncommon. The analysis of scent samples collected from identical twins living under the same environmental conditions and different environmental conditions will also be presented and compared. Various collection techniques for axillary scent and hand odor samples will be discussed as well as the stability of these samples when stored at room temperature. Sampling methods and mediums are also being investigated in an attempt to optimize the recovery and storage of human scent from forensic specimens. Different absorber types, including both sterile gauze and King’s Cotton, which is used by the Dutch National Police for human scent collection, have been evaluated based on their ability to collect human scent. Persistence/dissipation studies of human scent using SPMEGC/MS will also be presented evaluating variables such as light (UV, VIS) and temperature effects. Human Scent, Odor Profiles, SPME-GC/MS 111 B155 Identification of the Metal Residues on the Death Related to the High Voltage Spark Region by SEM-EDS Abdi Ozaslan, MD, Istanbul University, Department of Forensic Medicine, I.U. Adli Tip Anabilim Dali, Cerrahpasa, Istanbul, 34303, Turkey; Ismail Cakir, PhD*, Council of Forensic Medicine, Ministry of Justice, Adli Tip Kurumu Esekapi Cerrahpasa, Istanbul, 34246, Turkey; H. Bulent Uner, PhD, Istanbul University, The Institute of Forensic Sciences, I.U. Adli Tip Enstitusu Cerrahpasa, Istanbul, 34303, Turkey; Humman Sen, MD, Council of Forensic Medicine, Ministry of Justice, Turkey, Adli Tip Kurumu Esekapi Cerrahpasa, Istanbul, 34246, Turkey; Salih Cengiz, PhD, Istanbul University, The Institute of Forensic Sciences, I.U. Adli Tip Enstitusu Cerrahpasa, Istanbul, 34303, Turkey; and Sermet Koc, MD, Istanbul University, Medical Faculty of Cerrahpasa, I.U. Adli Tip Anabilim Dali, Cerrahpasa, Istanbul, 34303, Turkey Attendees will learn an easy method for SEM-EDS data from the area of metallic particles which were originated from the evaporating conductor contaminate the skin the arc-burns related to the high voltage. This presentation will impact the forensic community and/or humanity by demonstrating how SEM-EDS data from the area of metallic particles which were originated from the evaporating conductor can contaminate the skin the arc-burns related to the high voltage. High voltage electric energy wires, transformers, and the devices which run with this kind of electricity, may cause serious damage and death. It is the goal of this presentation to demonstrate this to those in forensic medicine by showing the conductive evaporated metallic particles existing on the arc-burn of the corpse using the scanning electron microscope-energy dispersive spectroscopy (SEM-EDS). It was reported to authorities that somebody who entered a transformer building in Istanbul without permission died of an arc which was formed by the high voltage within the building. The autopsy was performed in the Morgue Department of Council of Forensic Medicine, Ministry of Justice. During the autopsy, samples were collected using double-sided adhesive coated stubs from the surface of the skin lesions, which was the result of the high voltage arc, by stub. The removed samples were examined by SEM-EDS technique. The results were evaluated to determine the origin of death. It was concluded that the SEM-EDS data from the area of metallic particles originated from the evaporating conductor. Contaminating the skin, arc-burns resulting from the high voltage, are important combined with other close examinations to identify the cause and mechanism of death. Electrical Arc Burnt, Scanning Electron Microscopy-Energy Dispersive Spectroscopy, Metallic Particle B156 Development of a Method to Produce Lead Particle Patterns Howard A. Harris, PhD, JD*, and Abbey Scott, MS, University of New Haven, Forensic Science Program, 300 Boston Post Road, West Haven, CT 06516 Attendees will learn how to make reproducible lead oxide particle patterns of know amounts of lead oxide on various substrates. This presentation will impact the forensic community and/or humanity by assisting individuals interested developing improved methods of visualizing lead particle patterns and recovering GSR particles. 112 This research involves the development of a simple procedure to produce lead particle patterns on paper or cloth that resemble those used to estimate muzzle to target firing distances. Forensic examiners have long used the patterns produced on the target by lead particles and burned and partially burned smokeless powder particles to estimate the distance between the muzzle of a firearm and the object through which the bullet passed. Test patterns for evaluation of method variations were produced by firing a weapon from different distances into a piece of filter paper or cloth. This usually requires a trip to a firing range and produces rather variable patterns due to inconsistencies in the amount of smoke and lead produce by a weapon from shot to shot. In addition, it would be convenient to be able to measure the efficiency of various gunshot residue (GSR) recovery techniques. To be able to do valid comparisons it is necessary to be able to control the amount of lead deposited on the substrate. This is virtually impossible when producing patterns in the normal way at a firing range. The authors have tried to develop a laboratory method for producing patterns of fine lead oxide particles, similar to muzzle to target patterns, on paper and cloth substrates. One can purchase very fine lead oxide powders in the size range of GSR particles1. Suspending these particles in water, similar to the molybdenum sulfite suspensions used for the fingerprint reagent “small particle reagent”2, followed by delivering a known volume of the solution to the target would allow the delivery of a known amount of lead oxide to the target. A simple squirt gun can be used to produce fairly tight patterns similar to those produced at several inches by most firearms. To produce more diffuse patterns, similar to those observed from shots of one foot or greater distances, a simple spray bottle can be used. By weighing the squirt gun or spray bottle before and after a “squirt” one can calculate the amount of liquid delivered to the target. By knowing the concentration of the lead oxide in the suspension one can calculate the amount of lead being delivered to the target. Using varying concentrations of lead oxide in the suspension allows control of the amount delivered over a fairly large range. Three different substrates were used- filter paper, Whatman 3MM, Electrophoresis blotting pads S&S and white cotton cloth. It was found that using an inexpensive squirt gun fairly reproducible patterns could be generated by firing vertically down from a range of three to five feet. Using a spray bottle a more diffuse pattern could be made spraying from six inches to twelve inches in a horizontal direction. The patterns were visualized using the standard two step-sodium rhodizinate method3. As expected the diameter of the pattern increased as the distance increased. Similar patterns over a number of shots were reproduced. The patterns produced at the same distances on all three substrates were very similar in appearance. By weighing the squirt gun filled with distilled water it was determined that it deliver 0.40 g of water quite reproducibly. A lead stock solution containing 15g of lead oxide in 500 ml. of water was prepared. It was found that the patterns produced with this solution were too concentrated to resemble gunshot patterns. A working solution was prepared by diluting ten ml. of the stock solution to one hundred ml. This working solution was used to obtain patterns as indicated above and the patterns were visualized with sodium rhodizinate. This technique produced quite reproducible looking patterns the spread and intensity of which could be conveniently varied as indicated above. 1. Alfa Aesar Research Chemicals Iron Oxide -325 Mesh Powder 2. Advances in Fingerprint Technology, H.C. Lee & R.E. Geansslen, 2nd Ed., CRC Press, Chapter 4 p 113-4 3. M.R. Bartsh, H.J. Korbus, & K.P. Wainwrght, J. Forensic Sciences 1996;41: 1046-1051 GSR, Sodium Rhodizinate, Muzzle-Target Distance * Presenting Author B157 Lead Isotopes in Gunshot Residues Jurian A. Hoogewerff, PhD*, Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA, United Kingdom; and Herman Miesbauer, Ing, Upper Austria State Government, Goethestrasse 86, Linz, A-4021, Austria The goal of this presentation is to inform the forensic community about the possible usefulness of using lead isotope ratio analysis in specific cases. Different from the absolute concentration of an element or element/element ratios, radiogenic isotopic ratios from elements like lead or strontium are not sensitive to production or manufacturing processes and provided that no additional radiogenic isotope is added, the isotopic signature from the source material is retained. Due to geological processes on geological time scales most raw materials have developed identifiable isotopic fingerprints. Especially the lead isotopes are unique because the combination of three radiogenic isotopes (206Pb, 207Pb and 208Pb) and one stable isotope (204Pb) allow the construction of several different isotope ratio scatter diagrams which permit a very sensitive and discriminative assessment of the possible different sources of the lead. In a recent case the clothes of a murder suspect contained elevated traces of lead, possibly from GSR from a shooting incident. The defense argued that the lead contamination had been caused by exposure in a technical workshop. Additionally the evidence material had been kept under suboptimal conditions for several years that could not exclude contamination. Preliminary trace element analysis indicated elevated concentrations of several metals including lead. In this study the authors compared the lead isotopic composition of two GSR sheets from the murder victim with fifteen samples taken from a pullover of the suspect. The samples from the pullover were taken from the front and the back and the two sleeves in order to evaluate any focus of possible lead contamination from the GSR. This study showed that three different sources contributed to the lead found on the pullover but that none of these three sources agreed with the composition of the GSR found on the sheets. Lead Isotopes, Gunshot Residues, Isotope Analysis B158 The FBI Laboratory’s Response to Recommendations Regarding Comparative Bullet Lead Analysis Diana M. Wright, PhD*, and Marc A. LeBeau, MS, FBI Laboratory, Chemistry Unit, Room 4220, Quantico, VA 22135 Attendees will learn the FBI Laboratory’s response to recommendations regarding the analysis, assessment, and significance of comparative bullet lead examinations, which were offered in an independent report by the National Research Council of the National Academies. This presentation will impact the forensic community and/or humanity by informing the forensic community of the FBI Laboratory’s response to recommendations set forth in a report by the National Research Council of the National Academies regarding comparative bullet lead analysis and its interpretation. Comparative bullet lead analysis is the physical and chemical examination of lead bullets, fragments, or shot pellets. It is a non-routine examination, in that, it is only performed on damaged specimens that are unsuitable for direct comparison by a firearms examiner or in the absence of a firearm for comparative projectile testing. The FBI Laboratory has provided this examination for over 35 years in support of local and federal investigations involving recovered ammunition. Throughout the history of this examination, challenges have been raised in court as to its validity, scientific merit, and probative value. As * Presenting Author each criticism has been raised, the FBI Laboratory has sought to address these concerns through the use of publications and presentations at scientific meetings. Not all of the challenges, however, have concerned the scientific method employed. Questions regarding the significance of the examinations have consistently been posed to courts, which have chosen to allow the jury to decide the issue. Periodically, more comprehensive Daubert and Frye challenges have also been presented to the court regarding this examination. Comparative bullet lead analysis has successfully withstood each of these challenges. The FBI Laboratory has always welcomed constructive assessments of its scientific practices. The Scientific Working Group (SWG) program has been an excellent forum to allow for spirited scientific discussions that ultimately result in consensus documents to serve as sample protocols for the community. Unfortunately, comparative bullet lead analysis is an examination that is non-routine or non-existent in the FBI Laboratory’s peer organizations at both the state and federal level. Trace element analysis, in general, is still fairly non-routine for most laboratories. The necessary consumption of a portion of the evidence, the cost to purchase and maintain the requisite equipment, and the challenges associated with deriving a meaningful conclusion from the quantitative analysis of the composition of a mass– produced, man-made entity have all contributed to a lack of other laboratories embracing the technology. To provide an impartial assessment of the comparative bullet lead procedure, statistical analysis and subsequent testimonial assertions, the FBI Laboratory commissioned a study by the National Research Council of the National Academies as a means of addressing the challenges to this examination. The intent of this study was to obtain guidance as to what improvements might be necessary to continue to produce quality results that could be appropriately presented to a non-technical audience. To that end, the FBI Laboratory also requested that the NRC consider appropriate language to convey the bullet manufacturing process, chemical analysis, statistical assessments, and interpretative conclusions to a jury in a manner that was both thorough and concise. This presentation will summarize the committee’s recommendations to the FBI Laboratory. The recommendations cover a very broad range of topics including: the analytical protocol, the number of elements analyzed, quality control and proficiency testing measures, statistical recommendations for interpretation of the results, report wording, testimony language, and the use of phrases that the NRC committee developed to describe some of the concepts presented to a jury during direct testimony. The FBI Laboratory’s response to each suggestion will be described after presentation of the specific recommendation. Bullet Lead Analysis, NRC Recommendations, Comparative Examinations B159 Using LA-ICP-MS at the Netherlands Forensic Institute Shirly Montero, PhD*, Maarten Hordijk, Jan de Koeijer, Wim Wiarda, Ing, Peter de Joode, Ing, and Gerard van der Peijl, PhD, Netherlands Forensic Institute, Postbus 24044, Den Haag, 2490AA, The Netherlands Attendees will learn the discrimination potential of LA-ICP-MS for the elemental analysis of different types of materials of forensic interest. This presentation will impact the forensic community and/or humanity by demonstrating the discrimination potential of LA-ICP-MS in the analysis of chemical composition of some materials of forensic interest that cannot be discriminated using traditional methods. Trace evidence is widely recognized for its associative value. Sometimes recovered samples associated with a suspect and samples associated with a known source are compared to each other based on the physical properties as well as their chemical composition. The choice of methods to characterise and compare trace evidence depends on, among other factors, the accessibility of the proper instrumentation (i.e., sensi113 tivity, discrimination potential), the cost of the analysis including analysis time and the characteristics of the evidence (such as size and physical state). However, when there is control on the variation of physical properties in materials of the same kind, the discrimination potential of the methods used to measure such properties decreases. In many of those cases, elemental analysis has demonstrated to have a great value when used for the characterisation of such materials. In addition, if the sample is small, more sensitive methods are needed to measure reliably the elemental composition, in particular at trace levels. In recent years, different research laboratories and international networks have made an effort to develop and validate analytical methods with better sensitivity, precision and reproducibility. At the Netherlands Forensic Institute, different analytical methods have been optimised and validated for the elemental analysis of materials of forensic interest by LA-ICP-MS. Two of these materials, glass and pigment-based ink on paper, have been selected for this presentation. There are many studies demonstrating the value of elemental analysis of glass by different techniques. On the other hand, the need for non-traditional methods for the examination of documents produced with inkjet printers (e.g., threatening letters, contracts, and invoices) has increased only recently with the shifting from dye-based inks to pigment-based inks. Inks belonging to the latter type, are more robust when exposed to light and more resistant to solvents in contact with the paper, thus, not bleeding as much as inks of the former type. However, pigment-based inks are not soluble and therefore, not easily separated by traditional methods such as TLC and HPLC. The improved quality in the manufacturing of printers makes the differentiation among them based on physical characteristics more difficult, whether from the same manufacturer or from different ones. The technique, LA-ICP-MS, combines the sensitivity and precision of ICP-MS with the advantages of laser ablation sampling. With the use of LA-ICP-MS there is no need for laborious and lengthy digestion procedures with dangerous chemicals. In addition, the common analytical interferences that are increased by the presence of solvents are minimised using laser sampling, improving the detection limits of some potentially discriminating elements. The amount of material ablated for the complete analysis is very small (~fg), allowing the analysis (including replicates) of very small samples. The destruction is minimal with craters in the order of 10-8 m2, in contrast to the solution approach where the whole sample to be analysed is irreversibly digested. The system at the NFI is an ICP-MS (Perkin Elmer ELAN DRC plus) in combination with a 3 mJ- 213 nm-laser ablation system (New Wave UP-213). The standard reference materials (SRMs) NIST 612 and 1830 were used for the development and validation of a quantitative method for glass analysis. This method has been used to analyse glass samples collected by the police from different locations within the Netherlands. In addition, the traditional analyses such as the refractive index measurements and the less sensitive semi-quantitative µXRF measurements were carried out on all the samples. The methods, the selection of the matching criteria and the results of the comparisons of the samples using elemental concentrations and refractive index (measured with GRIM II) will be discussed. Finally, the development and validation and application of a semi quantitative method for the analysis of pigment-based inkjet inks will be also discussed. LA-ICP-MS, Elemental Analysis, Trace B160 Characterization of Trace Elements in Gunshot Residue of Lead Free Primers Winchester, Winclean, and Remington This presentation will impact the forensic community and/or humanity by demonstrating a systematic analysis, of gun shot residue (GSR) from primers and ammunitions in the area surrounding bullets holes and from the shooter’s hand. This is a very important tool to determine the shooting distance, types, of the primer and ammunitions to the forensic scientist. A comparative study of the gunshot residue particles produced during firing bullets having lead free primers manufactured by Winchester, Winclaen, and Remington were performed. Morphology and composition were analyzed using scanning electron microscope equipped with energy dispersive x-ray. The effect of lead pollution caused by the use of bullets and primers containing lead is one of the greatest problems facing the law enforcement and the military. Therefore attempts have been in progress to eliminate lead based primer mixtures and bullets. The recent development of lead free ammunition and bullets represent a new challenge for ballistic specialists and forensic scientist. Much research work has been carried out and published on lead based GSR but, only very few research studies have been reported in the analysis gunshot residues produced from lead free primers during firing. In this context this research is a study of the topography, morphology, and composition of the residues deposited during firing using lead free primers and ammunitions. Attempts have also been given to detect and estimate the amount of various constituents in the given ammunition and also to correlate the results to forensic examination. Test firing, collection of gunshot residues, and subsequent analysis were carried out at the Forensic Science Laboratory of the Albany State University. A specially made bullet trap filled with polymer fibers was employed for live firing. The firearms used in this study were a Springfield Armory 0.45 caliber (model 1911) and a Beretta 9mm Parabellum (model 92 FS.CaC). Two types of ammunition, 9 mm and 45 ACP, used in this study were manufactured by Winchester (Win Clean) and Remington. Firing distance and the area surrounding the target are the two parameters used in the collection of gun shot residues. A total of 60 rounds were fired in this study. Imaging and compositional analysis were performed on a Cam Scan 44 scanning electron microscope equipped with energy dispersive x- ray detector. Spherical and Non-spherical particles with size in the range 3- 30 m are studied for intensity, distribution and composition of gun shot residue particles. Intensity and distribution are studied as a function of shooting distance and collection areas. Number of particles on the shooter’s hand is high, compared to those collected from different areas of the target. The elements identified from the GSR particles of Winchester Winclean primers are Potassium, Calcium, Aluminum, Chlorine, Sodium, Copper, Zinc, and Silicon where as Remington constitutes Copper, Zinc, Magnesium, Chlorine, and Silicon. Composition variations are based on the size and shape of the GSR particles. Nonspherical and large particles always composed of more than three elements. Small and spherical particle contains either one or two elements. Attempts have been made to distinguish between Remington and Winchester Winclean GSR particles in terms of composition and morphology. Both types of GSR particles contain Copper and Zinc alone or in combination with other respective elements. This is because of the brass plated cartridge or the brass firing pin. Lead and barium is also found in both types due to use of previously fired gun. Morphology and size do not show any differences in their intensity or distribution. Characteristic GSR particles having spherical, spherical with dimples, oval, and irregular shapes are common in both types. Gun Shot Residue, Scanning Electron Microscope, Lead Free Primers Zachariah Oommen, MSc, PhD*, Scott Pierce, and Odister Kisha, Albany State University, 504 College Drive, Albany, GA 31705 Attendees will gain a comparative understanding of the gunshot residue particles produced during firing bullets having lead free primers manufactured by Winchester, Winclaen, and Remington. 114 * Presenting Author B161 Detection of Recent Handling of Firearms Joseph Almog, PhD*, The Hebrew University, Casali Institute of Applied Chemistry, Jerusalem, 91904, Israel; Yaniv Avissar, MSc, The Hebrew University, Analytical Chemistry Department, Jerusalem, 91904, Israel; Assaf Sagiv, MSc, The Hebrew University, School of Pharmacy, Jerusalem, 95744, Israel; and Daniel Mandler, PhD, The Hebrew University, Analytical Chemistry Department, Jerusalem, 91904, Israel Attendees will learn how to detect recent handling of firearms by a simple and sensitive color test. The reasons for differences between various individuals in the quality of the marks developed on their palms. This presentation will impact the forensic community and/or humanity by increasing the number of serious crimes, which are resolved by scientific techniques. Several physiological and environmental factors that affect the outcome of the Ferrotrace technique have been thoroughly studied. Consequently, two improvements, which greatly enhance sensitivity, have been suggested. Handling of a gun results in the formation of invisible impressions, caused by transfer of iron traces to the skin surface. Visualization of these impressions is possible by spraying the palms with a solution of 3-(2pyridyl)-5,6-diphenyl-1,2,4-triazine (PDT), which forms a magenta complex with iron(II) residues. Quantitative data are reported for the first time on the amounts of iron transferred to the hand upon holding a firearm. Iron levels between 21–315 ng/cm2 were found on volunteers’ palms after a single holding of a handgun. Determination of the iron traces was accomplished spectrophotometrically using PDT as a chelating agent. The transfer of iron from firearms to the palm was found to be, by and large, a chemical (dissolution) rather than mechanical dislodgement. The prime factor that determines the amount of iron transferred from the firearm to the hand is the moisture level on the palm. More factors, however, are involved in this process. Three time-dependent factors have been studied with relation to their effect on the developed mark: the gripping duration of the weapon; the time that elapses from the moment of contact; and the rate of iron dissolution in aqueous solutions containing sweat components in physiological concentrations. It was found that the amounts of iron transferred to the palm depend on both, the gripping period and the levels of palmar moisture. Thus, only a few seconds of gripping were required for good marks to develop (corresponding to 80 ng.cm-2 of iron) on highly-moistured hands (“good acceptors”). Much longer gripping periods were necessary for marks of similar intensity to develop on relatively dry hands (“poor acceptors”). Experiments aimed at studying the effect of sweat components on metallic iron dissolution were carried out in aqueous solutions. It was found that chloride ions in physiological concentrations remarkably enhanced the dissolution, while L-serine, the major amino acid in palmar sweat, had a detrimental effect on this process. These findings are likely to be of importance in courts of law, as well as in the war against terrorism and serious crime. Two modifications, which have been suggested − splitting the development process into two subsequent steps, and exposing the hand to water vapor − greatly improve the sensitivity of the method. Several cases that were resolved by the Ferrotrace reagent will be demonstrated. Forensic Science, Firearms, PDT * Presenting Author B162 A Portable X-Ray Fluorescence Instrument for Forensic Investigations Jeffrey Schweitzer, PhD*, University of Connecticut, Department of Physics, Unit 3046, 2152 Hillside Road, Storrs, CT 06269-3046; Jacob Trombka, PhD, and Samuel Floyd, NASA Goddard Space Flight Center, Laboratory for Extratraterrestrial Physics, Greenbelt, MD 20851; Carl M. Selavka, PhD*, Massachusetts State Police, Crime Laboratory System, 59 Horse Pond Road, Sudbury, MA 01507; Gerald Zeosky, MS, New York State Police, Forensic Investigation Center, 1220 Washington Street, Albany, NY 12226-3000; Norman Gahn, JD, Office of the District Attorney, Milwaukee County, Milwaukee, WI 54040; and Timothy McClanahan, BS, and Thomas Burbine, BS, NASA Goddard Space Flight Center, Greenbelt, MD 20851 After attending this presentation, attendees will understand the critical design features and operational parameters of a new, portable x-ray instrument for assisting in the recognition of trace evidence for crime scene and forensic laboratory applications. This presentation will impact the forensic community and/or humanity by describing the development of a portable x-ray fluorescence instrument; demonstrating the operational performance of the instrument for trace residua of forensic importance, including (but not limited to) primer residue, blood, and semen in a simulated crime scene; and discussing the future use of this instrument at crime scenes or in the laboratory to develop investigative leads by assisting in the recognition and recovery of such trace evidence through elemental analysis. A rugged x-ray fluorescence (XRF) instrument has been designed to investigate trace element content in and on evidence at crime scenes. The initial focus is to identify possible materials of interest such as gunshot residue and bodily fluids, but it has broad capability for general XRF applications. The instrument was designed to be part of a system to aid crime scene investigation and transmit the data to locations requiring it. This portable instrument was designed to meet the constraints of weight, battery operation, and ruggedness. Some special design features, however, were needed to achieve detection of microgram quantities of the trace elements of interest. This instrument is part of a Teleforensics program jointly funded by NIJ and NASA. This collaboration seeks to develop cost-effective instrumentation based on technology developed for the space program to benefit crime scene investigation, and to develop advanced instrumentation for planetary missions for NASA. A critical factor at crime scenes is the collection of evidence for analysis at forensic laboratories. The friable nature of evidence requires rapid recognition, to avoid losing the probative information contained therein. Some evidence is invisible to normal investigation techniques, either because it involves trace quantities not visible to any investigation technique, or because it is covered or hidden from view. Many types of potential evidence can be indicated by crime scene detection through in situ trace element analysis. Data is shown that supports possible use of a portable XRF instrument through detection of gunshot residue, blood (through the detection of the iron in hemoglobin), and semen (through the detection of zinc protoporphorin). To detect the low levels of trace element concentrations, advanced technology has been incorporated including the unique x-ray generator and a recently developed Shottky cadmium telluride x-ray detector. The design of the internal structure of the instrument is a critical component for minimizing the background due to coherent scattering. This design was necessary to ensure that the instrument could measure microgram quantities of elements of interest. It was also necessary to select the x-ray tube anode material to ensure that the tube’s x-ray line production does not interfere with the detection of elements of forensic interest. A key feature for the efficient and reliable operation of the instrument is spectral analysis software that can adapt to changing backgrounds and arbitrary elemental content. The results are incorporated in a database that was originally developed for planetary missions. In addition, it is now 115 understood that the application of historically-derived XRF data loaded into a relational database representing data relationships between items of interest and background matrices is just as important in forensic applications as has been recognized for planetary missions. Discussion of the use of such relational databases to assist in data recognition and recovery, and those data related to forensic trace evidence either available or researched to date, will also be included. The authors have also helped develop a Monte Carlo code for modeling the interaction of x-rays from any inputted source and the secondary particles that they produce as they interact with any type of material. This code can simulate the x-ray fluorescence produced by a variety of forensic materials (e.g., blood, semen, gunshot residue). Finally, results of experiments to date characterize the expected performance of the instrument for detecting trace element concentrations that are useful for investigating crime scenes and in laboratory applications. Teleforensics, Trace Recognition & Recovery, Portable X-Ray Fluorescence B163 Evaluation of SNPs as Tools in Human Identity Testing Peter M. Vallone, PhD*, Amy E. Decker, BS, Michael D. Coble, PhD, Margaret C. Kline, MS, and John M. Butler, PhD, National Institute of Standards and Technology, 100 Bureau Drive, Gaithersburg, MD 20899-8311 After attending this presentation, attendees will learn the utility of single nucleotide polymorphisms (SNPs) as human identity markers. The attendee will learn about different classes of SNP markers, typing technologies, and how SNPs compare with STR markers. This presentation will impact the forensic community and/or humanity by reporting on the utility of SNP markers to the DNA testing community. SNP markers can provide valuable complementary roles in human identity testing. The typing of coding region mtSNPs and small autosomal panels of SNPs for typing degraded DNA are two examples of where SNPs can benefit the forensic community. SNPs have the potential to play a helpful role in human identification testing. The small PCR amplicon sizes associated with SNP typing technologies make SNPs attractive for typing degraded DNA or other low copy number situations. SNP can be useful in combination with STRs for resolving complex paternity issues (e.g., incest), identifying victims of mass disasters where insufficient family references are available and possibly inferring population of origin. SNPs located in the coding region of the mitochondrial genome have been used to separate common HV1/HV2 mitotypes thereby extending the power of mtDNA testing (1,2). SNPs located on the Y chromosome have been evaluated for ethnicity prediction and individual sample discrimination (3). Various SNP typing platforms exist, but at this time there is not a universally agreed upon platform for SNPs and human identity testing. Currently researchers are typing SNPs with multiplex allele specific primer extension (ASPE) reactions. The assay is comprised of an initial step of PCR followed by primer extension and subsequent fragment separation and detection by capillary electrophoresis. ASPE multiplex panels can routinely type 6-12 SNPs in a single tube and have reported to go as high as 35 SNP markers. Important considerations for SNP markers are the larger number required to equal the discriminatory power compared to traditional STRs, their inability to resolve complex mixtures, issues related to databasing new loci, and the availability of a standard analysis platform. However, in appropriate situations SNPs can be useful as a supplementary tool complementary to STR markers. Methods and Materials: A total of 70 bi-allelic (C/T) SNP markers have been typed for 189 U.S. samples. Amplifications were performed in 6-plex panels. Amplicons between 59 –108 base pairs were generated. The 116 SNP markers were typed using multiplex ASPE assays and capillary electrophoresis. An 11-plex ASPE assay for typing coding region SNPs that helps resolve the most common Caucasian mitotype was developed and run on samples previously screened by Roche linear arrays for a common HV1/HV2 mitotype. Multiplex assays consisting of 3 ASPE multiplexes and 5 commercial hybridization multiplexes were used for typing 50 Y-SNPs. The 50 Y-SNPs were typed for 229 U.S. African American and Caucasian samples. Summary of Results: Novel multiplex SNP assays have been developed for typing various classes of SNP markers on U.S. population samples. Subset panels of the 70 autosomal SNP markers have been used to successfully type DNA from shed human hairs. Results for mtSNPs and Y-SNPs allow for an evaluation of the practical utility of various SNP markers in a human identity context. Samples typed by commercial and novel multiplex STR panels allow for a direct comparison of SNP and STR markers. Conclusions: Practical and inherent characteristics of SNP markers will prevent them from replacing traditional STR typing methods. However, SNP markers can provide valuable complementary roles in human identity testing. The typing of coding region mtSNPs and small autosomal panels of SNPs for typing degraded DNA are two excellent examples of where SNPs can benefit the forensic community. References: 1. Coble, M.D., Just, R.S., O’Callaghan, J.E., Letmanyi, I.H., Peterson, C.T., Irwin, J.A., Parsons, T.J. (2004) Single nucleotide polymorphisms over the entire mtDNA genome that increase the power of forensic testing in Caucasians. Int. J. Legal Med., 118: 137-146. 2. Vallone, P.M., Just, R.S., Coble, M.D., Butler, J.M., Parsons, T.J. (2004) A multiplex allele-specific primer extension assay for forensically informative SNPs distributed throughout the mitochondrial genome. Int. J. Legal Med., 118: 147-157. 3. Vallone, P.M. and Butler, J.M. (2004) Y-SNP typing of U.S. African American and Caucasian samples using allele-specific hybridization and primer extension. J. Forensic Sci. 49(4): 723-732. Single Nucleotide Polymorphism, Multiplex PCR, Degraded DNA B164 Allele Drop Out at Locus D5S818 Caused by a Single Nucleotide Polymorphism at a Primer Binding Site Peter Bugert, Institute of Transfusion Medicine and Immunology, Friedrich-Ebert Str.107, Mannheim, 68167, Germany; Lotte Henke, PhD*, and Juergen Henke, PhD, Institut fuer Blutgruppenforschung, Hohenzollernring 57, Cologne, 50501, Germany; and Harald Klueter, MD, Institute of Transfusion Medicine and Immunology, Friedrich-Ebert Str.107, Mannheim, 68167, Germany After attending this presentation, attendees will understand the reason for the loss of allele D5S818*10 when using PowerPlex®16 kit, approaches to overcome this and similar problems, and the impact and consequences for data bases. This presentation will impact the forensic community and/or humanity by providing the forensic community with the knowledge of the nature and the frequency of a primer binding site mutation and its impact on multiplex PCR analyses. Moreover, information is provided how to overcome the problem of allele drop out in general. The goal of this presentation is to clarify the reason for loss of allele *10 at locus D5S818 when using PowerPlex® 16 kit. Outcome: The authors present data about a single nucleotide polymorphism (SNP) at STR locus D5S818 including association of the mutant with the STR allele *10 and strategies to overcome the problem of allele drop out. Three SNPs were localized in close proximity to the D5S818 STR region at positions –13 (C/T), +4 (G/T) and +36 (T/C). The reverse primer * Presenting Author for amplification of this locus in the PowerPlex® 16 kit binds in the region of the +36T/C SNP. In order to investigate allele frequencies of the +36T/C SNP the authors established an allele-specific PCR method using sequencespecific primers (PCR-SSP). The PCR-SSP method was used for genotyping 288 samples from a DNA archive of blood donors from Southwest Germany. Four of the samples revealed a heterozygous genotype of the +36T/C SNP, thus, the frequency of the +36C-allele was 0.69% in this population. According to the Hardy-Weinberg-Equilibrium the heterozygous +36T/C genotype occurred at a frequency of 1.4%, whereas the homozygous +36C/C type may be observed in 1:20,000 individuals only. Sequence analysis of the D5S818 locus in the 4 individuals with the heterozygous +36T/C genotype demonstrated a linkage of the +36C-allele with the STR allele *10 in all cases. In the multiplex STR-analysis using PowerPlex® 16 kit the samples showed homozygous phenotypes at locus D5S818. The same samples were investigated further using the same primers for amplification of the D5S818 locus but in monoplex PCR. The monoplex analysis revealed heterozygous genotypes for all samples with unambigous amplification of allele *10 in addition to other alleles. In order to solve the problem of allele drop out in multiplex STR analysis the authors investigated the use of a PCR primer containing a wobble base at the corresponding position of the +36T/C SNP. The standard reverse primer for amplification of the D5S818 locus was replaced by the wobblebase primer in the multiplex assay. DNA samples with known STR- and +36T/C SNP-types were analyzed by using the modified primer mix. The STR allele *10 could be clearly detected even in samples with a heterozygous +36T/C genotype, indicating that the wobble primer is suitable to overcome the problem of allele drop out. On the other hand the modified primer led to an increased background, i.e., higher numbers of unspecific amplification products. Modification of the PCR program should overcome this problem. In cases of a homozygous D5S818 phenotype the authors would suggest the use of one of the following strategies: 1) Decreasing the annealing temperature; 2) Retyping of the samples using D5S818 monoplex PCR; 3) Genotyping of the +36T/C SNP using the PCR-SSP approach; 4) Use of a D5S818 wobble-base primer in multiplex STR-analysis. STR Locus D5S818, Aallele Drop Out, Single Nucleotide Polymorphism (SNP) B165 Some Interesting Point Mutations and Deletions Found Through STR Allele Sequencing sequencing results will be discussed in the context of the growing number of more than 230 variant alleles reported and cataloged as part of the National Institute of Standards and Technology STRBase website: http://www.cstl.nist.gov/biotech/strbase/. Methods and Materials: DNA sequencing primers lying outside of PCR amplification assay primer binding sites have been designed and tested for all 13 core STR loci used in the Combined DNA Index System (CODIS) as well as the D2S1338, D19S433, Penta D, and Penta E loci that are contained in commercial STR kits such as PowerPlex® 16, SGM Plus, and Identifiler. A variety of polyacrylamide gel electrophoresis conditions have been developed to separate closely spaced heterozygous alleles so that these alleles can be individually sequenced. Gel cutouts (individual alleles) are re-amplified prior to sequencing each allele. Variations in the individual alleles are determined by aligning their sequence to a reference sequence from GenBank. These alignments are assisted by the use of the software program Sequencher 4.1 (GeneCodes, Ann Arbor, MI). Summary of Results: The novel sequencing primers developed encompass the primer binding regions of all known published primer sequences for loci included in commercial STR kits and thus enable an examination of polymorphisms giving rise to allele dropout upon PCR amplification. Conclusions: Methodologies for DNA sequencing of STR alleles can aid in understanding the molecular basis for allele dropout due to point mutations or insertion/deletions in template DNA that disrupt PCR primer annealing. An increasing number of rare variant alleles are being discovered and information is being uncovered through DNA sequencing that can be helpful in assessing natural human variation and developing improved detection assays in the future. References: 1. Budowle, B., et al. (2001) STR primer concordance study. Forensic Sci. Int 124: 47-54. 2. Clayton T.M., et al. (2004) Primer binding site mutations affecting the typing of STR loci contained within the AMPFlSTR SGM Plus kit. Forensic Sci.Int. 139: 255-259. 3. Drabek, J., et al. (2004) Concordance study between miniplex STR assays and a commercial STR typing kit, J. Forensic Sci. 49(4): 859-860. Short Tandem Repeat DNA Typing, DNA Sequencing, Variant Allels B166 Loss of Heterozygosity at Several Loci of the 13 CODIS Core STR Loci in a Patient Diagnosed With Cancer Margaret C. Kline, MS*, Michael D. Coble, PhD, Jill E. Appleby, BS, Richard Schoske, PhD, and John M. Butler, PhD, National Institute of Standards and Technology, 100 Bureau Drive, MS 8311, Building 227, Room B250, Gaithersburg, MD 20899-8311 Katherine E. Long, BS*, Rick W. Staub, PhD, Judith I. Floyd, BS, and Robert C. Giles, PhD, Orchid Cellmark, 2600 Stemmons Freeway, Suite 133, Dallas, TX 75207 Attendees will learn a methodology for sequencing variant STR alleles will be described along with some interesting findings from samples producing allele dropout upon PCR amplification with various primer sets. This presentation will impact the forensic community and/or humanity by enhancing the knowledge of the forensic community in regards to methodologies used to define the differences found in variant alleles. Polymorphisms exist in the flanking regions of short tandem repeat (STR) loci that can cause allele dropout when they fall underneath PCR primer binding sites. The resulting “null alleles” are typically detected when concordance studies are performed using sets of PCR primers with different annealing positions (1,2). Some interesting deletions have been discovered in the flanking regions of D13S317 and VWA through concordance studies between miniSTR assays and commercial kits (3). In addition, several forensic laboratories have supplied samples possessing some novel variants that have been characterized. These STR typing and After attending this presentation, attendees will have an increased awareness of the possibility of a complete or partial loss of heterozygosity in a single source sample at multiple locations in nuclear DNA as a result of deletions due to cancer. This presentation will impact the forensic community and/or humanity by making scientists aware of the possibility of observing a loss of heterozygosity in a single source sample, which may be misidentified as a mixture when an inadequate number of loci are tested. It has been documented that a loss of heterozygosity in one of the systems of the 13 CODIS core STR loci can occur due to actual autosomal rearrangements associated with cancer. It is also known that a loss of heterozygosity can be observed in poor quality DNA samples from paraffin embedded tissues resulting in stochastic effects during amplification. In this particular case, a loss of heterozygosity was seen at more than one locus in several specimens with some specimens exhibiting a loss of heterozygosity at up to four loci. * Presenting Author 117 After a patient’s untimely demise from cancer, slides used in the patient’s diagnosis were submitted for DNA analysis via STR testing to ensure that the tissue on the slides was not contaminated by another patient’s tissue during the preparation process for evaluation by a pathologist. The patient in this case had initially been diagnosed with nonHodgkin’s Lymphoma. After treatment with chemotherapy, the lymphoma was declared to be in remission. Approximately one year later, the patient was found to have a growth in one of his lungs. This tumor was subsequently diagnosed as being small cell carcinoma also known as oat cell carcinoma. While being treated for the lung tumor, the patient suffered a relapse of the non-Hodgkin’s Lymphoma. Although it was noted that the lung tumor had begun to decrease in size the patient passed away a short time later. Upon autopsy, there was no indication of the lung tumor and the involved parties sent the paraffin blocks and related slides used to diagnose the small cell carcinoma to two private forensic DNA laboratories in order to determine the possibility of specimen mis-handling. Peak imbalances observed in the samples tested from the patient led the first forensics laboratory performing testing to conclude that the samples were a mixture of the patient and another individual. However, none of the eight loci tested exhibited more than two alleles. The second laboratory (Orchid Cellmark Dallas) then received three paraffin blocks and nine corresponding slides for testing and comparison with STR testing. Portions of the paraffin blocks and related slides were subjected to organic extraction via phenol chloroform and extracts were purified and concentrated using Microcon centrifugal filter units. The extracts were quantitated and initially amplified using Applied Biosytem’s Profiler Plus Kit. At a later date, the samples were also amplified using Applied Biosystem’s Cofiler Kit. After injection into Applied Biosystem’s 310 Genetic Analyzer, analysis was performed using Applied Biosysem’s GeneScan and Genotyper. This analysis revealed severe peak imbalances in several specimens at D3S1358 (in both Profiler Plus and Cofiler), Amelogenin (in both Profiler Plus and Cofiler), D8S1179, D21S11, D5S818, D13S317, D7S820 (in Profiler Plus only) and actual complete loss of heterozygosity at D5S818 and D13S317 in two samples. As seen in the data, the loss of heterozygosity was neither more prevalent in the smaller systems nor the larger systems. The peak imbalances varied from sample to sample inasmuch as there were not only differences in the percentages of peak RFU’s, there were also discrepancies as to which systems revealed peak imbalances. Throughout all thirteen loci, no more than two alleles were observed. Several months later, tissue slides prepared at autopsy were received and tested using methods identical to those used for the lung tissue slides. Although the autopsy slide tissue seemed to be more degraded than the lung tissue slides, the results obtained showed the same peak imbalances consistent with a loss of heterozygosity. Heterozygosity, STR, Cancer B167 Patterns of Allele Sharing in 13-Locus DNA Profiles of Siblings and Other Relatives Julie B. Kempton, MS, MFS*, S. Meredith Monroe, MS, Sarah A. Chenoweth, BA, Monica L. Ammann, MFS, Carter A. Cromartie, MFS, and Kristen Johnson, BS, Prince George’s County Police Department, DNA Laboratory, 7600 Barlowe Road, Palmer Park, MD 20785 After attending the presentation, the participants will gain an understanding of the usefulness and limitations of assessing whether similar DNA profiles may come from related individuals. Determining such relationships can be a valuable tool in conducting criminal investigations. This presentation will impact the forensic community and/or humanity by demonstrating Identifying the likelihood that DNA profiles 118 are from close relatives can be of great value to criminal investigations. This can aid investigators in identifying new suspects, and may result in faster apprehension of perpetrators of serious crime. In criminal investigations involving DNA evidence, it can be useful to know if DNA profiles may have come from related individuals. Previous studies have shown that the profiles of siblings match at six of the thirteen CODIS loci, on average. Since matches at five or six loci are not uncommon in unrelated individuals, the authors would like to assess whether two similar profiles are the result of coincidence or biological relationship. Profiles from over one hundred sets of siblings were compared, along with other first and second-degree relatives. The average number of identical loci (both alleles shared) in full siblings was 4.7, with a range of 1 – 9. The average number of non-matching loci (neither allele shared) was 1.5, with a range of 0-5. These results were compared with a database of several hundred unrelated individuals, with primary focus on those unrelated profiles with were identical at four or more loci. The average number of non-matching loci in these pairs was approximately 4, with a range of 19. Thus, the number of non-matching loci is the best predictor of whether two similar profiles are more likely from siblings than from unrelated individuals. However, because there is substantial overlap in the ranges, caution should be used when assessing the possible biological relationship of persons with similar DNA profiles. Another factor that should be considered is the overall expected frequency of the 13-locus profile. One individual in the database showed a high degree of similarity with a large number of unrelated profiles, matching at seven loci in two separate instances. The overall expected frequency of this profile is approximately 1 in 1 trillion (FBI Popstats). While this exceeds the threshhold for declaring identity, it is considerably more common than the frequencies observed for most 13-locus profiles. The results of this study have proved useful in several cases in the authors’ jurisdiction. In one such case, a bottle was obtained from the trash of a possible suspect in a series of sexual assaults. A 9-locus DNA profile obtained from the bottle matched the semen profile at six loci and shared one allele at the three other loci, a strong indication that the DNA on the bottle came from a sibling of the perpetrator (the suspect shared the house with two brothers). Based on this information, surveillance was increased on the suspect and a DNA sample was obtained when the suspect blew his nose and discarded the tissue on the sidewalk. This sample matched the semen profile from the sexual assaults, leading to the arrest and conviction of the rapist. DNA, STRs, Siblings B168 Application of Multiple Displacement Whole Genome Amplification to Forensic DNA Analysis Linda C. Rourke, MSFS, MPhil*, Annette T. Lee, PhD, and Peter K. Gregersen, MD, North Shore/LIJ Research Institute, Department of Genomics and Human Genetics, 350 Community Drive, Manhasset, NY 11030; and Margaret Wallace, PhD, and Peter R. De Forest, DCrim, John Jay College of Criminal Justice, 445 West 59th Street, New York, NY 10019 The goal of this presentation is to convey research findings evaluating the fidelity of whole genome amplification by multiple displacement amplification from DNA extracted from aged bloodstains. This presentation will impact the forensic community and/or humanity by describing research on the forensic applicability of a technique which may allow forensic scientists to overcome the problem of limited sample sizes and open the door for many additional types of forensic DNA analysis that can be useful in the investigation of crime. The recent completion of the human genome sequencing project has created many opportunities for further elucidation of human genetics. Such * Presenting Author advances continue to fuel the explosive growth of forensic DNA technology. The bottleneck in forensic science is that the quantity and condition of forensic evidence samples frequently limit the extent of analysis. Recent improvements in whole genome amplification may allow forensic scientists to overcome the problem of limited sample sizes and open the door for many additional types of forensic DNA analysis that can be useful in the investigation of crime. Multiple displacement amplification (MDA) is a method of whole genome amplification (WGA) which is capable of providing large quantities of human genomic DNA from limited sample sources. It is a relatively new method which, although validated for various types of clinical samples (whole blood, cheek cells, etc.), has yet to be tested on the type of samples commonly encountered as forensic evidence. This research seeks to determine whether WGA/MDA is a suitable means of amplifying human genomic DNA from samples of a forensic nature. Current forensic DNA methods amplify targeted areas of the human genome in order to produce a genetic profile for the sample of interest. However, forensic evidence samples are frequently limited to that which is collected from a crime scene. A method that could accurately and representatively amplify the entire sample genome would provide sufficient DNA for multiple analyses and for archival storage. Minute samples could be amplified to ensure a sufficient quantity of DNA for both prosecution and defense teams to analyze. Amplification of the entire genome would also create sufficient DNA to analyze genomic loci that provide information useful in determining phenotypic characteristics of the unknown source of biological evidence from a crime scene. This research will determine whether WGA/MDA can provide accurate STR profiles from DNA extracted from dried bloodstains. Blood will be drawn from anonymous donors for the preparation of control DNA and bloodstains. Control DNA will be extracted from fresh blood. Known volumes of blood will be stained onto a white cotton substrate, dried, and stored at room temperature. Stains will range in size based on the original volume of fresh blood used to stain the substrate: 1µl, 10 µl, and 50 µl. As an extraction control, the same volumes of blood will be aliquoted directly into microfuge tubes and stored under the same conditions as the bloodstained swatches. DNA will be extracted from the stains and extraction controls at various time points, ranging from Day Zero to 10 weeks, using Gentra’s PureGene kit. Extracted DNA will be quantitated using PicoGreen fluorescence. DNA extracted from aged bloodstains and extraction controls will be amplified using Amersham’s GenomiPhi Kit. Control DNA from the freshly drawn blood will also be amplified to compare the aged versus control DNA. The amplified DNA will be quantitated using PicoGreen fluorescence. The whole genome amplified DNA will then be analyzed for 7 of the 13 short tandem repeat (STR) loci required by the FBI for inclusion in the CODIS database system. The seven STR markers are D5S818, D13S317, D7S820, FGA, D3S1358, D21S11, and D18S51. Control DNA that has not been amplified via GenomiPhi will also be analyzed in these 7 loci. STR profiles from control DNA will be compared to the STR profiles obtained from the GenomiPhi amplified aged bloodstain samples to determine if the latter samples are capable of providing accurate DNA profiles. Forensic DNA Analysis, Multiple Displacement Amplification, Whole Genome Amplification * Presenting Author B169 Degenerate Oligonucleotide-Primed PCR: ‘Proofreading’ a Method for Forensic DNA Analysis Denise N. Rodier, BS, Virginia Commonwealth University, 1000 West Cary Street, PO Box 842012, Richmond, VA 23284; Kristen M. Meyer, MFS, North Carolina State Bureau of Investigation, 121 East Tryon Road, Raleigh, NC 27603; and Kristen E. Lewis, BS, and Tracey Dawson Cruz, PhD*, Virginia Commonwealth University, 1000 West Cary Street, PO Box 842012, Richmond, VA 23284 The goal of this research project is to provide the forensic DNA community with a Whole Genome Amplification (WGA) tool – Degenerate Oligonucleotide-Primed PCR (DOP-PCR) – that can readily increase the success rate of analysis of degraded, aged, or otherwise compromised biological evidence samples using existing conventional lab technologies and standard procedures for data analysis. This presentation will aim to build upon the data that was previously presented by showing improved amplification success using a dual-enzyme approach. This presentation will impact the forensic community and/or humanity by allowing for an increased success rate for cases where a very low amount or unusable DNA was obtained from a crime scene, either through lack of evidence altogether or through severe degradation of samples. These samples can be analyzed by technicians without additional training or equipment, and with minimal additional costs to the labs. Whole genome amplification techniques have been utilized in a number of molecular diagnostic areas, including embryonics, cancer biology, histopathology, and in a variety of studies on molecular diseases and genetic linkage. Degenerate Oligonucleotide-Primed PCR (DOPPCR) is one such whole genome amplification technique which allows for low copy number DNA samples to be preamplified such that high quality, high yield DNA samples can be available for downstream forensic STR amplification and analysis. Thus far, standard DOP-PCR techniques have shown to sufficiently generate enough high yield, high molecular weight DNA for STR analysis by capillary electrophoresis. However, results show some preferential amplification and allele/locus drop out in these subsequent multiplex STR amplifications. By adding a proofreading enzyme (i.e., Pyrococcus furiosus) in a small ratio to the Thermus aquaticus enzyme currently used with DOP-PCR and increasing extension times during thermalcycling, longer-sized products should be achieved. This will greatly reduce the preferential amplification and allele/locus drop out seen in multiplex STR amplifications. Initially, input DNA amounts ranging from 0.25 nanograms to 7.5 picograms were tested in four DOP-PCR setups using either Taq enzyme with Taq buffer, Pfu enzyme with Pfu buffer, 16:1 Taq/Pfu enzyme combination in Taq buffer, or 16:1 Taq/Pfu enzyme with Pfu buffer in either 50ul or 100ul reaction volumes. All resulting DNA was visualized by agarose gel electrophoresis and human DNA was quantitated by the traditional Quantiblot method. Preliminary results indicate that while the standard DOP-PCR reaction (using Taq enzyme in Taq buffer) produced products with a size range of approximately 250bp to 2,000bp, the Taq/Pfu enzyme combination produced products with a size range of approximately 500bp to over 5,000bp. Additionally, preliminary data from the Taq/Pfu enzyme combination experiments indicate that input DNA amounts as low as 7.5 picograms (~2 cells) yielded enough DNA to be visualized on a 1% agarose gel, and DNA yields increased by several thousand-fold for samples with as low as 62 picograms input DNA. Further research will evaluate the ability of DOPPCR products produced with the enzyme combination approach to generate a correct profile with distinct and balanced peaks at all multiplexed STR loci with minimal stochastic variation. In addition, other DOP-PCR enzyme combinations and WGA techniques will also be evaluated in a similar manner. 119 These research findings will impact the forensic DNA community by allowing for an increased success rate for cases where a very low amount or unusable DNA was obtained from a crime scene, either through lack of evidence altogether or through severe degradation of samples. These samples can be analyzed by technicians without additional training or equipment, and with minimal additional costs to the labs. Whole Genome Amplification, DNA, STR Analysis B170 Assessment and In Vitro Repair of Damaged DNA Templates Ashley M. Hall, MS*, and Jack Ballantyne, PhD, University of Central Florida, National Center for Forensic Science, PO Box 162366, Orlando, FL 32816-2366 Attendees will learn methods for the assessment and repair of damaged DNA templates derived from forensically relevant samples. At the present time, little is known about the damage done to the DNA in biological stains exposed to various exogenous insults. This presentation will impact the forensic community and/or humanity by presenting the first comprehensive study of this damage at the molecular level and intoduce methods for the repair of the damaged DNA, facilitating the recovery of a genetic profile. DNA extracted from biological stains is often intractable to analysis. This may due to a number of factors including a low copy number (LCN) of starting molecules, the presence of soluble inhibitors or damaged DNA templates. Remedies may be available to the forensic scientist to deal with LCN templates and soluble inhibitors but none presently exist for damaged DNA. In fact, knowledge of the biochemical nature and the extent of DNA damage in physiological stains is rudimentary at best. Also unknown is the point at which the damage inflicted upon a particular sample precludes the ability to obtain a genetic profile for purposes of identification. Therefore, the primary aims of this work were first ascertain the types of DNA damage encountered in forensically relevant stains, correlating the occurrence this damage with the partial or total loss of a genotype, and then to attempt the repair of the damage by means of in vitro DNA repair systems. The initial focus of the work was the detection of damage caused by exogenous, environmental sources, including factors such as UV irradiation, heat, and humidity. By incorporating various lesion specific enzymes, a set of assays, both PCR and gel-based, have been developed which describe the type and extent of damage inflicted upon DNA, both in a hydrated and dehydrated state. Using these procedures, the major causes of damage have been identified and their effects on genetic profiling assessed. Armed with this knowledge, the next focus was the repair of the damage by means of in vitro DNA systems. Efforts have been concentrated on base excision repair, single strand gap repair, and translesion synthesis assays. By modifying the assays and employing various combinations of the systems, a genetic profile has been obtained from previously intractable samples. DNA Damage, In Vitro DNA Repair, Lesion Specific Endonucleases B171 Age Determination: The Identification of Newborns Using Messenger RNA Profiling Analysis Michelle Alvarez, BS*, and Jack Ballantyne, PhD, University of Central Florida, Department of Chemistry, PO Box 162366, Orlando, FL 32826-2366 Attendees will learn a method to determine if a bloodstain originated from a newborn baby. This presentation will impact the forensic community and/or humanity by demonstrating the development of a novel method, which can be incorporated into the forensic laboratory, to aid in investigations concerning bloodstains thought to originate from newborns. It is now a matter of routine for the forensic scientist to obtain the genetic profile of an individual from DNA recovered from a biological stain deposited at a crime scene. Potential contributors of the stain must either be known to investigators (i.e., a developed suspect) or the questioned profile must be searched against a database of DNA profiles such as those maintained in the CODIS National DNA database. However, in those instances where there is no developed suspect as yet or there is no match with any database sample, the DNA profile per se presently provides no meaningful information to investigators, with the notable exception of gender determination. To aid in these investigations another useful biometric that could provide important probative information is the age of an individual. For example, the ability to provide investigators with information as to whether a DNA donor is a newborn baby, an adolescent teenager or an elderly individual could be useful in certain cases, particularly those involving young children such as kidnapping or in providing additional intelligence during terrorist investigations. Currently no reliable validated molecular tests are available for age determination. The lifecycle of humans comprises a number of developmentally recognized stages. As the human proceeds through these developmental stages, sub-sets of the 30-50 thousand human genes will be differentially expressed. Theoretically, and given sufficient knowledge of developmental genetics, a determination of the global gene expression profile could reveal constellations of genes whose expression is correlated with a specific age. One example of how developmental regulation of gene expression can lead to the determination of age is by examining the ß-hemoglobin locus located on the short arm of chromosome 11 (11p15.5). This chromosomal region encodes five functional ß-like globin genes, ε, γG, γA, δ and ß, each with a specific pattern of highly regulated developmental gene expression. In the first weeks of neonatal development embryonic hemoglobin (ξ2, ε2) is produced by the yolk sac. Around twelve weeks of gestation embryonic hemoglobin synthesis decreases and the fetal liver, spleen, and bone marrow begin producing fetal hemoglobin (α2, γ2), which continues throughout fetal development. Shortly after birth fetal hemoglobin production decreases and the synthesis of adult hemoglobin (α2, ß2) rises and is the major form of hemoglobin present throughout life. Therefore, the development of an assay which selectively identifies an increased presence of fetal hemoglobin (γ chain) in a biological stain, would infer that the donor of the stain is a newborn baby. Descrived here are two novel methods which assay the age related levels of variant forms of gamma hemoglobin (HBGv) present in bloodstains, with an increased level of expression being indicative of the newborn status of the individual. The first method is a duplex reverse transcription-polymerase chain reaction (RT-PCR) and allows for the determination of a newborn based on a present / absent gamma hemoglobin product, along with the presence of an internal control, the ribosomal protein S15. A second realtime-PCR (qPCR) assay, can distinguish between newborns and other age groups by evaluating the ∆Ct values (Ct S15 - Ct HBG) generated. A positive ∆Ct value would indicate that a bloodstain originated from a newborn; in contrast a negative ∆Ct value would be obtained with all other ages. Age Determination, Identification of Newborns, Messenger RNA Profiling 120 * Presenting Author B172 mRNA Profiling: Body Fluid Identification Using Multiplex Real-Time PCR Jane S. Juusola, BS*, University of Central Florida, Department of Chemistry, PO Box 162366, Orlando, FL 32816-2366; Kevin W.P. Miller, PhD, FBI Academy, Building 12, CTFSRU, Quantico, VA 22135; and Jack Ballantyne, PhD, University of Central Florida, Department of Chemistry, PO Box 162366, Orlando, FL 32816-2366 Attendees will learn a novel means of identifying body fluids of forensic interest. This presentation will impact the forensic community and/or humanity by demonstrating the mRNA based approach, such as the multiplex real-time PCR method described here, possibly allowing for the facile identification of body fluid stains and is one of many assay platforms that could conceivably supplant the battery of serological and biochemical tests currently employed in the forensic serology laboratory. Conventional methods of body fluid identification use labor-intensive, technologically diverse techniques that are performed in a series, not parallel manner, and are costly in terms of time and sample. Furthermore, for some frequently encountered body fluids no confirmatory technique exists. There is no definitive test, for example, for the presence of saliva or vaginal secretions. In seeking to develop novel multiplex (i.e., parallel) analysis procedures for body fluid identification that are compatible with current DNA analysis procedures, The authors have chosen assays based upon messenger RNA (mRNA) since it is expressed in a tissue type specific manner. Terminally differentiated cells, such as blood lymphocytes, ejaculated spermatozoa, or epithelial cells lining the oral cavity, have a unique pattern of gene expression, which is evinced by the presence and relative abundance of specific mRNA species. If the type and abundance of mRNAs can be determined in a stain or tissue sample recovered at the crime scene, it would be possible to definitively identify the tissue or body fluid in question. Advantages of an mRNA-based approach, compared to conventional biochemical analysis, include greater specificity, simultaneous, and semi-automated analysis though a common assay format, improved timeliness, decreased sample consumption, and compatibility with DNA extraction methodologies. It was previously reported that it is possible to isolate total RNA of sufficient quality and quantity from biological stains to enable subsequent detection of particular mRNA species using the reverse transcription-polymerase chain reaction (RT-PCR) technique and that has identified candidate sets of blood-, saliva-, semen-, vaginal secretions-, and menstrual blood-specific genes using a combination of literature and public database searches. In the present work, the authors report the development of a set of multiplex real-time PCR assays for the definitive identification of blood, saliva, semen, vaginal secretions, and menstrual blood. Real-time PCR employs a 5' nuclease assay to detect specific amplimers and eliminates the need for post-PCR processing and gel electrophoresis. The real-time instrument is capable of multi-color detection, and so by using probes labeled with different reporter fluorophores, it is possible to develop multiplex assays for body fluid identification. Real-time PCR also has the ability to quantitate target sequences, which is important in establishing the tissue-specificity of a gene product, particularly when the relative abundance of a number of different mRNAs can demonstrate a unique or restricted pattern of expression. Real-time PCR triplexes were developed that are composed of two body fluid-specific genes and one housekeeping gene and have been optimized for the detection of blood, saliva, semen, vaginal secretions, and menstrual blood as single or mixed stains. The methodology is based upon determining the delta Ct (∆Ct) values generated using the Ct for the housekeeping gene (HSK) and the Ct for each of the body fluid-specific genes (BFG) (Ct HSK- Ct BFG). Depending upon the body fluid-specific gene being tested, a positive ∆Ct value would indicate the presence of a particular body fluid, while a negative ∆Ct value would indicate the absence of that body fluid. * Presenting Author An mRNA based approach, such as the multiplex real-time PCR method described above, could allow the facile identification of the tissue components present in a body fluid stain and is one of many assay platforms that conceivably could supplant the battery of serological and biochemical tests currently employed in the forensic serology laboratory. mRNA Profiling, Multiplex Real-Time PCR, Body Fluid Identification B173 Individualization of Acer rubrum Using Amplified Fragment Length Polymorphism Margaret M. Wallace, PhD*, and Christine Bless, BS, John Jay College of Criminal Justice, Department of Sciences, Room 4510, 445 West 59th Street, New York, NY 10019; and Heather Palmeter, BS, America's Choice Healthplans, 1429 North 11 Street, King of Prussia, PA 19406 Attendees will learn how the AFLP technique is a rapid, inexpensive, and efficient method that can be used to genotype DNA from botanical material. Within the closed set of samples used in this study, the AFLP profiles were species specific and unique. AFLP is a rapid, cost-effective, simple, and robust method that can be used to genotype DNA of any origin and complexity. The technique can be used by forensic scientists to type DNA from nonhuman biological evidentiary material (plants, insects, and animals). This presentation will impact the forensic community and/or humanity by adding to the repertoire of techniques available to forensic scientists and increase the probative value of nonhuman DNA evidence. Amplified fragment length polymorphism (AFLP) is a powerful method that combines techniques from classical hybridization-based and PCR-based genotyping strategies. AFLP was used to genotype leaf material from red maple trees, Acer rubrum, which are widely distributed throughout much of the United States and Canada. Duplicate samples from 40 Acer rubrum trees were collected from four different sites in central New Jersey. Samples from five additional species were collected for comparison. One set Acer rubrum samples was used to establish a DNA database. The second set of Acer rubrum samples and the comparison samples were analyzed and compared to the AFLP profiles in the database to determine the discriminative capacity of the technique. The AFLP protocol was performed using components from the AFLP Core Reagent and Preamp Primer Mix I kits (Invitrogen, Rockville, MD). Genomic DNA was double-digested by two restriction endonucleases: EcoRI and MseI. The DNA fragments were ligated to EcoRI and MseI oligonucleotide adapters to generate primer binding sites. In this manner a select set of DNA fragments can be amplified without knowledge of the sequence. Two consecutive PCR reactions (preamplification and selective amplification) were performed. In the preamplification reaction DNA fragments were amplified using primers complementary to the adapters and adjacent restriction sites with one selective nucleotide at the 3' ends: EcoRI (5'-GACTGCGTACCAATTCA-3'; MseI (5'- GATGAGTCCTGAGTAAC-3'). During selective amplification the products of the preamplification reaction were amplified using a primer pair with three selective nucleotides at the 3' ends: EcoRI [5'- (dyeD4) GACTGCGTACCAATTCACT-3', Proligo, Boulder, CO] labeled with the fluorophore (D4WellRed, Beckman Coulter); MseI (5'- GATGAGTCCTGAGTAACAT-3', Invitrogen). The primer design and amplification strategy ensured that only a subset of the EcoRI- MseI fragments was preferentially amplified. The DNA fragments were separated by capillary electrophoresis using the CEQ 8000 DNA Fragment Analyzer. Data were analyzed using the CEQ 2000XL (Beckman Coulter) and Twin Peaks (authors, proprietary) software. Within this closed set, AFLP profiles were species specific and unique. The individualization of plant matter will enable forensic scientists to derive more information from evidentiary material and to help link a suspect or a victim to a particular crime scene (source tree). DNA, AFLP, Acer rubrum 121 B174 Extraction and Amplification of Nuclear DNA From Shed Dog Hairs Colinette S. Heath, MSc*, Eleanor A.M. Graham, MSc, and Guy N. Rutty, MD, MBBS, FRCPath, University of Leicester, Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester, Leicestershire LE, United Kingdom Attendees will learn how to successfully extract and amplifly several highly variable canine microsatellites for unique identity testing from shed dog (Canis familiaris). Upon completion of this research DNA profiles of unrelated dogs from different breed origin will be produced. This will not only provide verification of identity but will also display genetic diversity within inbred dogs. This presentation will impact the forensic community and/or humanity by providing a valuable asset to forensic science by opening new possibilities for linking suspects to crime scenes or victims. Hypothesis: That it is possible to extract genomic DNA from shed dog hairs to facilitate the animal's identification, parentage verification, and possibly link suspects to crime scenes or victims. At present, very little effort has been made to extract nuclear DNA from dog's hair and in previous studies this has often been proven unsuccessful. This study, however, provides information on the retrieval of genomic DNA from shed dog hairs and has shown that though difficult it is indeed possible. Text: DNA profiling of STR loci is now well established and extensively used for human identity testing such as identification of missing persons, investigation of crimes, cases of disputed paternity, investigation of mass disaster, and solving historic cases. However, the process of individual genetic identification has not been limited to human; attention has been paid to other organisms, including farm animals and pets. In addition to parentage verification and identification, nuclear and mitochondrial DNA profiling of animal samples (blood, saliva, tissues, and hairs) has contributed to homicide investigations and convictions. Shed animal hair is one of the most common biological materials recovered during forensic evidence collection. Extraction and successful PCR amplification of DNA from recovered hair could provide powerful intelligence to aid criminal investigations. Previous studies have however, shown that shed hairs contain only minute amounts of undegraded DNA, therefore analysis of nuclear DNA is mostly unsuccessful. This paper however, presents results of genetic profiling of nuclear DNA extracted from naturally shed dog hairs. Experimentation with three different extraction methods was undertaken, a modified Chelex extraction method proved the most successful in extracting amplifiable nuclear DNA. Modifications of the Chelex extraction include pre-extraction preparation with proteinase K, incubations at 56°C and 100°C plus micro-concentration of the solution. The quantity of extracted nuclear DNA was shown to be adequate for PCR based typing at 3 loci. This study shows that nuclear DNA can reproducibly be obtained from shed dog hairs. Dog Hair, DNA Extraction, DNA Profiling 122 * Presenting Author Engineering Sciences C1 Injury Pattern Analysis in Fatal Traffic Crash Investigation Michael D. Freeman, PhD, DC, MPH*, Oregon Health and Science University School of Medicine, 3071 Dogwood Drive South, Salem, OR 97302; and Clifford Nelson, MD, Medical Examiner Division, Oregon State Police, 301 Northeast Knott Street, Portland, OR 97212-3092 After attending this presentation, attendees will understand some principles of crash investigation, the necessary elements for the application of Injury Pattern Analysis, characteristic injury patterns of certain types of crashes, and an example of a practical application of Injury Pattern Analysis This presentation will impact the forensic community and/or humanity by serving as a key aspect of fatal crash investigation as it can augment traditional means of investigation in a systematized format via interdisciplinary communication and collaboration. Reconstruction of a fatal crash can be augmented, in certain circumstances, by information gleaned from the postmortem evaluation. Further improvement of the scope and accuracy of an investigation can result from evaluation of the injuries of crash survivors, taking into account the conformity of individual vehicle interiors as well as the movement of the occupants during the crash. The term “Injury Pattern Analysis (IPA)” is proposed as a description of a fatal crash investigation technique that utilizes accident investigation, and reconstruction techniques, occupant kinematics, postmortem records, hospital and healthcare provider acute injury records, and other evidence as an adjunct to the investigation of homicides resulting from fatal crashes. The authors present a case study in IPA as an example of the practical application of the technique. It is recommended that medicolegal death investigators become familiar with the principles of IPA. Crash, Fatal, Investigation C2 Numerical Models in Motor Vehicle Accident Reconstruction With Case Studies Matthew Weber, BSE*, Chris Van Ee, PhD, Design Research Engineering, 46475 DeSoto Ct., Novi, MI, 48377; and Boyd Allen, MASc, PE*, MacInnis Engineering Associates Inc., 21076 Bake Parkway, Suite 106, Lake Forest, CA, 92630 The goal of this presentation is to equip the attendee with an understanding of which collisions are suitable for numerical simulation techniques and some of the sensitive and insensitive input parameters used in the simulations. This presentation will impact the forensic community and/or humanity by making the forensic community aware to highlight sensitive input parameters in commercial accident reconstruction simulation software and contrasts some of the available software. This presentation demonstrates that a variety of computer programs are capable of generating similar and accurate results. Reconstruction of a motor vehicle accident using different computer software packages renders similar results if the packages are used appropriately. However, pre and post impact driver actions/reactions need to be interpreted with caution and can affect the results of the analysis. High-speed collisions can be analyzed using several available tools, which are more directly applicable than for low speed collisions. This is due in part to the fact that much of accident reconstruction theory and available test data originated with test speeds in the 30-35 mph range. A brief overview of the types of tools available for reconstructing high-speed accidents is presented. Several popular computer programs employing these tools and the associated assumptions are utilized in case studies. A * Presenting Author high-speed crash test in which one vehicle is driven into the side of a stationary vehicle and an actual “real world” left turning accident with two moving vehicles are used for illustration and comparison of the available methods and computer packages. Several computer programs, which are widely used for accident reconstruction, are used to reconstruct the collisions. The programs WinSlam© and EDCRASH™ are used to analyze the impacts from an energy standpoint. The programs PC Crash, EDSMAC4™, and DyMESH® simulate the impact and subsequent postimpact travel. The results of the various methods and their relationships to the known crash test parameters will be addressed. Additionally, some of the assumptions and inputs required by the tools are varied to illustrate their effects on the final results. Some simulation programs are not always directly applicable to low velocity impacts. “Low velocity” impacts between motor vehicles is a somewhat arbitrary term that here will refer to collisions involving very little or no physical crushing of the vehicles. Low velocity collisions often cannot be adequately analyzed using some traditional high-speed numerical techniques. Accident Reconstruction, Computer Modeling, Simulation Comparison C3 Weakness in the Numerical Models Used in Accident Reconstruction Programs John J Smith, MSEE, MSBMT*, Raymond P. Smith and Associates, 43766 Buckskin Road, Parker, CO 80138 After attending this presentation, attendees will understand the limitations of using computer programs in accident reconstruction. This presentation will impact the forensic community and/or humanity by identifying the limitation of the computer programs allowing the forensic community to understand when the results provided are significant and when they have little value. In the last several years, there has been an increasing usage of equations from various programs, like CRASH (Calspan Reconstruction of Accident Speeds on the Highway) and SMAC (Simulation Medel of Automobile Collisions), their derivatives, and other programs to determine pre impact and post impact velocities of vehicles. The use of equations from various programs, without regard for the underlying simplifying assumptions and checking routines, leads to errors and inaccuracies. The underlying physics of accident reconstruction has been analyzed and compared to the assumption used in several programs. The first area addressed were those based on CRASH. The analysis was performed by comparing the simplifying assumptions in the programs to the actual values expected in collisions. Additionally the simplifying assumptions were checked for internal consistency. One area of particular concern was if the program violated a simplifying assumption required in the derivation of the equations necessary to develop the program. As an example, if the underlying equations used a simplifying assumption of homogeneity and then the program instructed the user to input values that established the surface was not homogeneous, this was identified as a potential problem. Finally, the computer-generated results of tests were compared with the actual speeds of the vehicles to determine if the program was accurate A fundamental principle of mathematics is that for every unknown in a problem, a separate, independent equation must exist in order to arrive at a unique solution. In accident reconstruction the common unknowns of interest include: the mass of Vehicle 1, the mass of Vehicle 2, the initial speed of Vehicle 1, the initial speed of Vehicle 2, the final speed of Vehicle 1, the final speed of Vehicle 2, the approach angle of Vehicle 1, the approach angle of Vehicle 2, the departure angle of Vehicle 1 and the departure angle of Vehicle 2. 123 Less common, although often-critical variables include, tire forces, friction, steer angles, stiffness values, slope, surface material, and tire design. In order to resolve the problem, equations are often solved simultaneously. Common fundamental equations used are conservation of linear momentum, conservation of angular momentum, conservation of energy, the principle of restitution, Newton’s Laws of Motion, and the basic equations of motion. As the complexity of the problem increases, more equations are required to achieve a unique solution. As the number of variables and equations increase, the use of computers becomes more beneficial. This, in turn, explains the proliferation of programs available. Even the use of computers does not relieve the investigator of the basic need of a separate equation for each variable. Compounding the problem is the use of quadratics, in some of the equations, since quadratics usually do not have a unique solution. For this reason, it is common to use simplifying assumptions and secondary equations such as those postulated in the work of Campbell or McHenry. Often these secondary equations are based on the same or additional assumptions. As a result of the analysis it was determined that the numerical models used to predict the impact speed of vehicles have several limitations. The underlying simplifying assumptions used to derive the equations of the CRASH model in particular were found to have numerous problems. Among the critical assumptions identified in the derivation of the algorithm were several that were immediately violated by the program. These assumptions included that the vehicles act like a mass with a spring, the spring constant is constant, only plastic deformation occurs, the spring constants for both vehicles are equal, crush is symmetrical on both vehicles, the crush distance equals the acceleration distance and the system acts like a simple harmonic oscillator. Occasionally some of these assumptions are met, but it is rare that all are met and it is common that none are met. Typically, the program requires the operator to violate most, if not all of these assumptions. In addition to the primary part of the program, the crush coefficients used are derived by a separate approach that uses many of the above assumptions and actually adds additional ones including the assumption that the vehicles are homogenous both vertically and horizontally. The full-scale crash test run to validate the model actually showed that the results had no statistical significance. In some cases the program returns impossible answers. In other cases, the error associated with CRASH predictions has been observed to exceed 100%. The use of CRASH, and its derivative programs, to reconstruct automobile collisions is valid only under certain conditions. The results obtained have limited statistically value. Anytime the programs are used, the assumptions should be checked for validity and how well the assumptions are met by the facts of the collision. The approach used in this analysis has application in evaluating the reliability of any program used in accident reconstruction. Computer Model, Accident Reconstruction, Crash C4 Understanding Injury Potential in Low Damage Automobile Collisions Peter Alexander, PhD*, Raymond Smith & Associates, 43766 Buckskin Road, Parker, CO 80138 The goal of this presentation is to provide further insight into correlation between vehicle damage and impact severity. Observation of body damage coupled with frame deformation may provide criteria by which to judge impact speeds in situations where a visual examination of the vehicle shows little or no damage. This presentation will impact the forensic community and/or humanity by describing impact speed thresholds, which can prove helpful in understanding the injury severity in what otherwise, might appear to be a very low speed impact. INTRODUCTION: Raymond Smith and Associates is sometimes confronted by cases in which a vehicle has been impacted by another 124 vehicle, resulting in little or no visible damage. The driver of the target vehicle has sustained some fairly serious injuries, as determined by their physicians. How can the significant occupant injuries be reconciled with the apparent lack of significant vehicle damage? An approach has been developed, which demonstrates that in a number of these cases, despite the absence of obvious visible vehicle damage, the frame (or unibody) of the target vehicle was deformed by the force of the collision. A number of recent cases involve collisions in which a vehicle’s driver or passenger sustained significant head, neck, and back injuries. A physician diagnosed the injuries. The vehicle inspection often showed little to no obvious visible damage. Sometimes the bumper cover was scuffed, or there was a slight indentation in the bumper cover. Detailed body shop repair estimates for the vehicles, when available, showed either no damage or a need to replace only the bumper cover. A reconstruction expert claimed that because the damage level was so low, the bullet vehicle impact speed was under 5 m.p.h. and the speed change of the injured party’s target vehicle was less than 3 m.p.h. Thus it was unlikely that they would have been injured. THE APPROACH: When examining the target vehicle, the author looked at the fit of the hood, the trunk lid, and the doors. The eye can easily see a distortion of less than 1 millimeter in the fit of these items. If the spacing between these components and the vehicle’s body appeared to be uneven, a precision frame measurement was recommended. In every one of 14 recent cases, where visual evidence of external body distortion was observed, the precision frame measurement verified that the frame or unibody had been deformed beyond factory specifications. The 14 cases shown in Table 1, involved different vehicles impacted from the front, the rear, and the sides. In most of the cases where this method was applied, the frame or unibody distortion was approximately 1 centimeter. This is not necessarily sufficient to cause difficulty steering, or uneven tire wear. It is, however, indicative of a very forceful impact at a speed far above the 5 m.p.h. cited by the other expert. If no evidence of frame distortion was observed, removal of the bumper cover and disassembly of the bumper system often revealed deformation of the underlying bumper structure. Following the collision, in several cases, the bumper cover popped back out to its pre-impact position, masking the damage. Frame deformations of ~1 centimeter are indicative of impact speeds in the range 15 to 25 m.p.h. This speed range can be identified because, in a number of cases where frame distortion was observed, the impact speed could be independently determined based on post impact movement of the vehicles involved. Bumper damage generally does not appear in vehicleto-vehicle collisions below 8 to 10 m.p.h. Frame distortion is generally not seen below 12 to 15 m.p.h. At impact speeds in excess of 25 m.p.h. one would expect to see serious body damage to the vehicles. The literature contains reports of numerical modeling efforts, which attempt to correlate the forces imposed on a vehicle, in a collision, with the expected deformation of the vehicle. At present, these models are still in their infancy, and are not capable of predicting the type of deformations, which are the subject of this paper. At some point it may be possible to determine the impact speed, which caused a particular frame deformation, using a finite element numerical modeling approach. UNDERSTANDING INJURY POTENTIAL: The technical literature supports the view that there is no minimum or threshold vehicle impact speed (or speed change) required for occupant injury to occur. Logically, the higher the speed change, the greater the probability of injury. A portion of the impact energy is channeled into the vehicle’s occupants. Injury to the occupants can occur as a result of the acceleration forces and from interaction between the occupant and the interior components of the vehicle, including the seat belt. Observation of body damage coupled with frame deformation may provide criteria by which to judge impact speeds in situations where a visual examination of the vehicle shows little or no damage. The impact speed vs. damage described can prove helpful in understanding the injury severity in what otherwise might appear to be a very low speed impact. * Presenting Author Table 1 FRAME/UNIBODY DEFORMATION CASE TABULATION IMPACTED VEHICLE IMPACT LOCATION IMPACT SPEED (mph) REPAIR $ DEFORMATION (mm) INJURY 1992 Nissan Maxima Left Side, L.F. Tire 25-35 Tire Involvement 4691 4 Lumbar, required spinal fusion 1995 Honda Accord Left Front End 25-35 Tire Involvement 6805 6 No data 1997 Ford F150 PU Right Rear 37-40 1954 7 Ruptured L5-S1 1994 Dodge Ram 1500 PU Rear 15-25 3367 14 Herniation L5-S1 1992 Buick Le Sabre Rear >25 736 32 Stenosis at L3/4 , L4/5 and L5/S1 2000 Ford E250 Van Front & Rear Moderate 3839 5 1995 Ford Explorer 2WD Rear 20-35 4031 18 1998 Toyota Corolla Rear 15-25 422 6 1992 Ford T.Bird Rear End 20-25 5354 11 Herniated disk at L5 2002 Saab Left side, front tire 20-35 - 38 Cracked teeth, back problems 1989 Ford Tempo Rear 22 ~500 11 Herniated disks C4-C5 1991 Chevrolet Blazer Rear End** 20-35 2888 8 3 back surgeries: laminectomy, fusion 1998 GMC Suburban Rear End 15-25 110 8 Closed head injury, cervical injuries 1999 Jeep Wrangler Rear End 20-25 None 17 TMJ, Back injuries Brain injury Herniation L4/5 and L5/S1 Required spinal fusion Injury, Collision, Frame C5 A Proposed Practice for the Correct Forensic Interrogation of Non-Volatile Memory Data in Evidentiary Vehicle Electronic Control Units William Rosenbluth, MS*, Automotive Systems Analysis, Inc., 12015 Canter Lane, Reston, VA 20191 This presentation will impact the forensic community and/or humanity by discussing information on new forensic practices and procedures in a new and growing area of forensic technical investigation, electronic data recovery and documentation. It is important for the practicing forensic professional be familiar with proper procedures so that he or she can avoid pitfalls or errors that can jeopardize forensic findings and Court credibility, because existing practices and procedures often do not cover methods of electronic data recovery and documentation. SYNOPSIS: This presentation discusses methods and considerations for the examination and interrogation of non-volatile memory data in evidentiary vehicle electronic control units (ECUs) that may have been involved in an event or incident that may be reasonably expected to be the subject of litigation. This practice is intended to become applicable when it is determined that examination or interpretation of such non-volatile memory data may produce probative evidence. These methods are being considered for a new Practice for the Investigation of Non-Volatile Memory Data in Evidentiary Vehicle Electronic Control Units, currently being considered by ASTM committee E30.05 on Forensic Engineering. LEARNING OBJECTIVES: By observing the elements of this proposed protocol, the attendee will learn several methods for conducting an examination for the controlled interrogation of the data in a subject vehicle * Presenting Author electronic control unit (ECU). Such data may be saved in ROM, EPROM, EEPROM1 or flash-memory, all of which are non-volatile forms of memory. The retrieval of such data is commonly referred to as a download of information from the subject device2. A further objective of this proposed protocol is to retrieve any such data with the highest assurance of not changing or disturbing that data, either by erasure or overwriting. Such a download is referred to as a forensically neutral3 download. A forensically neutral download can be accomplished with a load box/interface containing proper electrical loads and interfaces, or by using an exemplar vehicle for the same purpose. An example of a non- forensically neutral download is shown in Figure 1, and an example of a forensically neutral download is shown in Figure 2. Figure 1 Figure 2 Certain commonly used commercial interrogating tools are not forensically neutral when used in a direct umbilical mode to interrogate SRS ECUs (i.e., a direct connection to the SRS ECU). In that mode, certain external fault codes will be added (or re-detected) because there is no provision for proper dummy-load resistors in the tool cabling. If the data of interest is not changed (e.g., crash data parameters), then a non-forensically-neutral interrogation may be acceptable. If certain fault codes are of 125 interest, then a non-forensically-neutral interrogation may not be acceptable. If a potential DTC data change is not acceptable, other test equipment, laboratory breadboards and/or the use of an exemplar vehicle can avoid this problem. In general, it is expected that the test conductor will have a proper test fixture, and a proper exemplar component to demonstrate that his/her test bed is forensically neutral. If the subject component is considered as evidence in litigation, and the manufacturer or supplier of the subject ECU is, or may be, a party to that litigation, it shall be considered standard practice to give the opportunity of first (qualified, non-destructive, non-intrusive, non-altering) download to the manufacturer or supplier of the ECU, with all adverse parties observing. This does not apply to exploratory interrogations or analyses of ECU components, which may be considered exemplar components. THEORY OF THE ANALYSIS: Electronic data within any ECU is an encoded representation of information, constants and variables used to govern the function of an electronic control unit, as well as to describe and document the version and level of such data. Such data are normally saved in a binary bit format. In order to create a concise representation of such data, it is usually represented by eight binary bit values, and these values are commonly represented as bytes of data, each byte having a specific memory address. Although most of the data referenced in this practice are saved in EEPROM, certain other data can be saved in ROM, EPROM or flash memory. In general, the test conductor should perform two test series, with two devices under test (DUT). The first series should involve an exemplar device (to provide a baseline verification of the test fixture) and the second series should involve the subject ECU. A. Power off test fixture (can include optional use of subject/exemplar vehicle as test fixture). B. Select DUT. C. Install DUT. DUT installation should include physical stability integrity if there is any chance that physical movement during the interrogation cycle will change the data within the DUT. D. Power on test fixture with DUT installed. Observe MIL codes and other-indicator status. Visually record. E. As appropriate, interrogate DUT with standard scan tool to observe scanner data (DTCs & PIDs). Save data as appropriate (photographically, computer data file, hardcopy, etc.) F. Interrogate DUT with interrogation tool to download EEPROM and/or selected PID information (RAM, ROM, EAROM, EEPROM). Save data as appropriate (photographically, computer data file, hardcopy, etc.) G. Re-interrogate with standard scan tool to record scanner data (DTCs & PIDs). Save data as appropriate (photographically, computer data file, hardcopy, etc.) H. If no exceptions, power down and remove DUT. I. Select next DUT as applicable and repeat steps 1-8. (For subject DUT, repeat EEPROM download procedure twice.) J. End test operations. 1 EEPROM = Electrically Erasable Programmable Read-Only Memory. EEPROM is made by using a special semiconductor construction that allows it to retain previously stored data even when the battery is disconnected. Flash-memory has similar characteristics to EEPROM, and that technology is also commonly used to save music or digital camera images in other commercial devices. 2 Common usage identifies “download” as the process of interrogating an on-vehicle ECU and recording that data on an external diagnostic ECU (laptop computer), and that common usage is preserved herein. When requesting PID information that usage is reasonably applicable. However, strictly speaking, SAE J2190:4.23 identifies that the process of requesting the transfer of data from an on-vehicle ECU to an external ECU (Mode $35) is called an upload request, whereas the process of requesting the transfer of data from an external ECU to an on-vehicle ECU to an (Mode $34) is called an download request. Again, to avoid confusion, common usage is preserved herein. 126 3 Not all electronic data interrogation processes are forensically neutral. A forensically neutral interrogation is one that will neither add or subtract error codes (DTCs) or crash data information from any ECU under interrogation. This applies to in-vehicle and bench top interrogation processes, including ECUs interrogated via direct umbilicals while still mounted in a vehicle. To be forensically neutral, such a unit must include provisions for actuator (squib, solenoid, etc.) dummy loads, sensor detection loads, MIL loads, serial feedback or for seatbelt switch status, so that any ECU undergoing such test-via-umbilical-interrogation will see only a correct operating environment during power-on and continuous loop checks. Electronic Data Recorders, Forensic Neutrality, EDR Protocol C6 The Need for Adequate Strength Seat Back Design - A Case Study Kurt D. Weiss, MSME*, Automotive Safety Research, Inc., 5350 Hollister Avenue, Suite D, Santa Barbara, CA 93111-2326 After attending this presentation, attendees will understand an analysis of an automobile collision during which a rear seat occupant was ejected while wearing a 3-point lap and shoulder belt. This presentation will impact the forensic community and/or humanity by demonstrating how an inadequate seat back design, while certified by federal safety standards, failed and resulted in catastrophic injuries THEORY OF THE ANALYSIS: Seat back strength is essential in order to provide adequate protection and containment of occupants in passenger vehicles. Seat back failure as a result of rear-end collisions reduces the potential for occupant restraint and increases the risk of occupant ejection. Federal Motor Vehicle Safety Standard (FMVSS) 207 establishes the requirements for seat assemblies and their attachment hardware to minimize the possibility of component failure by occupant forces acting on them as a result of vehicle impact. However, real world rear-end impacts have shown that inadequate strength seat back designs can lead to lack of restraint and ejection of vehicle occupants. COLLISION OVERVIEW: On the evening of October 6, 2002, a young male driver was operating his 2000 model year passenger vehicle at high-speed. Onboard were passengers in the right front seat and right rear seat. As the vehicle entered a downhill right-hand turn, the driver lost control and the vehicle began to yaw counter-clockwise. The vehicle veered left, crossing the opposing traffic lane, and struck a guardrail at the roadway edge. The initial guardrail strike was to the vehicle’s right front corner. The principal direction of force (PDOF) of this impact was approximately 90 right of center. As a result of the right front impact, the vehicle continued to rotate counter-clockwise and struck the guardrail at the right rear corner. Rotating through this right rear corner impact, the rear of the vehicle then impacted a guardrail post. The impact to the right rear corner along with the rear guardrail post impact had a combined change in velocity of approximately 12.1m/s with a PDOF of 150 right of center. After separating from the guardrail, the vehicle continued to rotate counterclockwise until it rolled to a stop on the shoulder. FORENSIC ANALYSIS: Inspection of the seat belts revealed forensic evidence confirming that all three passengers were wearing their 3-point lap and shoulder belts at the time of impact. In addition, photographs taken by investigating officers show the right rear seat belt still fastened at the collision scene. The right rear seat back was deformed due to occupant loading during the rear impact. In response to this impact and subsequent vehicle rotation, the right rear occupant ramped up the seat back, sliding under the seat belt, and was ejected through the rear hatchback window. The ejected occupant sustained fatal head injuries. The autopsy report revealed this occupant suffered an atlanto-occipital disarticulation, brainstem laceration and contusion, multiple skull fractures, and bilateral cerebral subdural and subarachnoid hemorrhage. * Presenting Author The subject vehicle rear seat back is a 50/50 split-back design such that either half can be independently unlatched and folded forward to increase cargo space. The latching mechanism for the left and right seat back panels are located outboard on the interior of the vehicle quarter panels. There is no restraining device or reinforcing structure at the centerline of the upper area of the seat back, except for a narrow fiberboard shelf behind the seat that conceals the rear cargo area when the hatchback is closed. FMVSS 207 section 4.3.2.2 specifies the acceleration requirements for the restraining devices of hinged or folding seat backs of forward-facing seats. The section requires that once engaged, the device shall not release or fail when subjected to a horizontal acceleration of 20gs opposite to the direction the seat back folds. Section 5.1.2 of the standard specifies the testing procedure such that the horizontal force is applied through the CG of the seat back panel. The right rear seat back panel has a mass of approximately 4.5kg: therefore, at 20gs a total force of approximately 890N would be applied. Based on the location of the latching mechanism and hinge point of the seat back panel, the 890N-applied force would subject the latching mechanism to a force of approximately 672N and the hinge point to a force of approximately 218N. A simple haversine analysis was used to determine the peak acceleration of the vehicle as a result of the rear impact. Assuming a collision pulse duration of 100ms, the resulting peak acceleration was approximately 24.6g. Medical records indicate the right rear occupant had a mass of 115kg, approximately 14 percent greater than the mass of a Hybrid III 95th percentile male test dummy. The mass and CG location of the individual torso and upper body components of a 95th percentile ATD were used to approximate the CG height of the effective mass of the right rear occupant. The effective mass, approximately 70% of the overall mass, is that portion of the occupant’s mass that loaded the seat back panel in the rear impact. The CG height of the effective mass is approximately 30.5cm above the seat cushion surface. Based on this analysis, the potential peak force applied to the latching mechanism was approximately 9,200N, and the potential peak force applied to the hinge point was approximately 8,400N. The potential peak forces applied to the latching mechanism and hinge point in the subject collision far exceeded the forces required by the seat back panel restraining devices to satisfy FMVSS 207. However, these calculated potential peak forces are not unlike those forces reasonably anticipated in real world collisions of this magnitude. Clearly, the seat back panel and its restraining devices were inadequately designed resulting in catastrophic failure in this unfortunate collision event. CONCLUSION: Seat back yield strength is an important consideration in foldable rear seat back designs. Anticipated occupant loading forces resulting from real world collisions far exceed the static force requirements used to certify seat back designs. Inadequate strength restraining devices can lead to seat back failure and increase the risk of occupant ejection. Seat Back Strength, FMVSS 207, Occupant Loading C7 Go-Cart Fatality, Engineering Case Analysis Adam K. Alexander, PhD, PE*, Aleksander & Associates, PA, PO Box 140558 Boise ID 83714, 5109 North Sawyer Avenue, Boise, ID 83714 The goal of this presentation is to present a typical product liability case which illustrates many aspects of engineering design for consumer products, and considers the special requirements and duty of care when the consumer market includes children. This presentation will impact the forensic community and/or humanity by demonstrating the complete cycle of consumer product design, marketing, consumer injury (fatality), litigation, engineering investigation, trial, jury verdict, and government (CPSC) response to remedy the product by recall. All too often, the process is short circuited by settlements, without addressing the possible latent hazards in consumer products. * Presenting Author Summary/Outcome: This paper presents the findings of an investigation into the circumstances of a go-cart fatality. A 13-year-old girl was driving a go-cart when her long hair was caught in the rotating machinery behind the seat. The case (Woodburn v Manco) went to trial, and the plaintiff’s verdict resulted in a recall of the go-carts. The presentation will include exhibits from the investigation and trial, with data and commentary. This typical product liability case illustrates many aspects of engineering design for consumer products, and considers the special requirements and duty of care when the consumer market includes children. The young driver was presented the cart, her brother’s, and instructed in its use. She was provided with her father’s motorcycle helmet, and tucked her long braided hair into her T-shirt, and then proceeded to drive the go-cart in a completely fenced dirt corral. The tire tracks show regular large circles, about 75 feet in diameter in an unobstructed venue. Her brother, nearby, investigated when he heard the engine idling for a prolonged time. Preliminary work included the examination of coroner’s photos that were taken at twilight and included a cover over the victim. Photogrammetric techniques were used on an exemplar go-cart to determine the victim’s actual position in the go-cart. An autopsy was not performed, and the cause of death was attributed to a neck fracture at C1C2. Further photogrammetric studies helped determine the likely length of the victim’s hair prior to the accident. A key element of this case was to establish the likely seating position of the victim, and the range of positions that the foreseeable go-cart user population may assume. Exemplars of wigs were used in conjunction with a subject pool of young and older male and female drivers of varying statures. The wig enabled a study of the location of the long braided pigtail, both in a tucked-in location in the driver’s T-shirt, and in the free position, dangling behind the go-cart seat. A further study illustrated the foreseeability of long hair in the general population in both male and female drivers. The evidence presented showed that the go-cart industry and their component suppliers had a longstanding knowledge of prior scalping incidents involving long scarves, long hair, and other clothing. However, the testing by the manufacturer and remedial actions were cursory, incomplete, and ineffective. These included a recall based on a rear axle entanglement accident. One of the consequences of the prior cases was the attempt by the manufacturer to warn the user by means of a label, and instructions. These Human Factors engineering related issues proved to be inadequate, given the well understood hierarchy of remedial engineering safety actions, namely that a warning is not an adequate resolution of a known hazard when a design change can eliminate the hazard. One of the engineering aspects of this case was the demonstration that a simple guard, at a nominal cost was sufficient to eliminate the nip point, the physical root cause of this fatality. Indeed, after this accident, the go-cart industry adopted a safety standard that incorporated just such a guard. The accident itself was linked to the speed of the vehicle, largely unknown since there were no witnesses at the time. However, a simple analysis suggests that even at moderate to slow speed the entanglement occurred in fractions of a second. The neck fracture was thus almost instantaneous. An opposing expert argued that a proper fitting helmet would have somehow ameliorated the situation, while another offered the opinion that based on his questionnaires, the girl was operating the vehicle in an unsafe manner, and that her parents were negligent in letting her drive. This was part of the defense strategy to allocate blame to the plaintiffs, Idaho being a comparative fault State, wherein a 50% negligence finding negates any award in favor of the plaintiffs. The verdict of the jury trial found in favor of the plaintiff’s, but the defense succeeded in persuading the jury to find 50% contributory negligence thus negating the damage amount. On the basis of the verdict however, the Consumer Products safety Commission (CPSC) directed a product recall in August of 2000 for 91,000 Go-Carts manufactured by Manco Products Inc., of Fort Wayne, Ind. The jury verdict regarding the negligence was appealed to the Idaho State Supreme Court, but did not prevail. Go-Cart, Human Factors, Anthropometry 127 C8 Momentum and Elastic-Plastic Vehicle Collisions: A Daubert Accuracy Analysis Richard W. McLay, PhD*, 1231 Hamilton Court, Iowa City, IA 52245; and Donald J. Anderson, PE, Anderson Engineering, 8504 Cottagewood Terrace, Blaine, MN 55434 The goal of this presentation is to present to the forensic engineering community an analysis of the validity of a momentum and elastic-plastic model for vehicle collisions to examine the Daubert error requirement. This presentation will impact the forensic community and/or humanity by examining the theory of three methods for predicting velocity in accident reconstruction. It makes clear that the method using pre- and post-impact motion in comparison with the highway witness marks is the most accurate. As such, it provides a means for guaranteeing the civil rights of defendants in criminal actions. This paper will present an analysis of the right-angle impact between two 4-wheeled passenger vehicles that are defined by their dimensions, masses, moments of inertia, and vehicle crush. The equations of mechanics in impact are integrated using a Taylor’s expansion of the impact force, proving that the error in the mean force is proportional to the time of impact if the first term in the series is defined by the impulse between the vehicles. The equations are also shown to be invariant with respect to the impulse on first integration. The second integration to yield the displacements at impact is shown to have meaning only if measurable crush is found on the vehicles. By using an elastic-plastic model for crush and demonstrating that the elastic motion is dominated by the plastic crush, closed-form expressions are derived for both the time-of-impact and the mean force of impact. The error analysis for the Daubert requirement on accuracy continues with the developments from the equations of mechanics. By using the time-of-impact derived from the crush, the angular changes of a target vehicle of 4105 lbs (1866 kg), moment of inertia of 2285 lb sec ft2, and bullet vehicles between 1500 lbs (682 kg) and 6000 lbs (2727 kg), shown in the figure below, were found. The results for angle changes are shown to yield an error in angle less than 5o that produces a cosine of 0.996 and a sin of 0.087. This proof demonstrates that impulsive motion theory can be used to study pre- and post-impact motion, validating the requirement of Daubert on knowledge of error. By using a standard ASTM tensile test for a body part of an American car, the theory of plasticity is shown to closely approximate a perfectly plastic steel model, at least for moderate strains near the yield point. The error induced by ignoring strain hardening is estimated and the resulting errors in approach and departure velocities are determined for the common pair of vehicles in the example. Finally, the chaos in crash testing of vehicles to determine the coefficients of crush is examined relative to the requirements of Daubert. In this study, the literature has been searched to examine multiple crash tests of identical vehicles in identical crashes. The theory of chaos is shown to apply to these crash tests because the deformation of the vehicle in crush is dependent on small changes in the initial conditions of the crash in both the structural geometry and in the angles of approach. Chaos appears in the results because these small changes in initial conditions cause disproportionately large changes in the folding patterns of the vehicle panels and 128 beam supports. As is predicted by the theory of chaos, derived crush coefficients have considerable variation. Even with a large sample size, there can be no guarantee that a given crash will follow one family of crush parameters over another. It would seem that the finite element method would yield the best results, assuming that the vehicle is available so that the crush sequence can be defined and measured. This makes the analysis a deterministic model. The closure of the paper describes the validation of the theory of the basic equations of mechanics through the study of impulse-momentum. It cautions the accident reconstructionist to make sure that predictions for the motions of the vehicle come from both the witness marks on the highway (using photogrammetry) as well as any crush and momentum simulation attempted. Of particular concern is the use of such impact models to determine speed when it is known that chaos is present in the crash tests from which crush data are determined. The authors’ conclusion is that it is better to use the equations of dynamics to study the highway witness marks than to depend on v calculations of a chaotic system. Impact, Momentum, Chaos C9 How Prevalent are Defective Automobile Air Bags? Peter K. Alexander, PhD, PE*, Raymond Smith & Associates, 43766 Buckskin Road, Parker, CO 80138 The goal of this presentation is to address questions regarding the reliability of automobile air bags. This presentation will impact the forensic community and/or humanity by answering questions regarding the reliability of automobile air bags. INTRODUCTION: The automobile air bag is just one part in a complex electronic and mechanical system designed to inflate the air bag at the proper moment. Air bag malfunctions can manifest themselves in four ways. The air bag can deploy for no reason, the air bag can deploy early or late, the air bag can inflate only partially when full inflation is called for, or the air bag can fail to deploy when it should have. This paper addresses only the last issue. Raymond Smith and Associates is currently working on 4 separate cases in which air bags failed to deploy. A reconstruction of the collisions showed that in all these cases, the air bags should have deployed. This surprisingly large number of failure to deploy cases raises the following question. Are there a large number of defective air bag systems in the current vehicle population? CRITERIA FOR AIR BAG DEPLOYMENT: In general the air bag should deploy in a frontal or side impact if the speed change induced by the crash is between at least 15 m.p.h. The air bag should never deploy at a speed change of less than 9 m.p.h. The speed change required for deployment with some of the newer two stage airbags and smart air bag systems that identify whether the occupant is small and sitting close to the air bag, can be higher. CRITERIA FOR AIR BAG NON DEPLOYMENT TO MANIFEST ITSELF: Three things have to happen before an air bag nondeployment event will come to public attention. 1) The air bag system must be faulty 2) An accident must take place, where deployment should have occurred but did not 3) An injury sufficient to raise the visibility level of the failure, occurred The air bag system contains sensors, a microprocessor, the air bag charge, the air bag, and a significant quantity of electronic circuitry. Due to the complexity of the system, there can be a large number of reasons for the air bag not to deploy when it should have deployed. Among the reasons the system might fail to deploy are the items that follow: * Presenting Author • An open circuit or communication failure within the system • A short circuit within the system • Sensor drift or improper sensor calibration • Voltage drift in the power circuitry • Software malfunction • A microprocessor malfunction • A faulty air bag charge • Environmental issues, such as moisture or contaminant intrusion during production or while in use • Poor system maintenance (an error code is displayed, indicating a problem, but no repair is performed) THE MAGNITUDE OF THE PROBLEM: A sense of the seriousness of the air bag failure to deploy problem can be gained from examination of the National Highway Traffic Safety Administration’s National Accident Sampling System Data. Every year the U.S. Government examines, analyzes, and documents key elements of a sampling of U.S. vehicle crashes. Most years the sampling contains data from 4,200 to 4,400 crashes. Selecting from the NASS database one and two vehicle crashes, which involved frontal impacts resulting in a speed change in excess of 15 m.p.h. produced the data shown in Table 1. This vehicle speed change should have caused the air bags, in air bag equipped vehicles, to deploy. Table 1 Air Bag Deployment and Non Deployment Collisions for 2 Injury Levels YEAR -Injury Severity COLLISIONS COLLISIONS WHERE AIR WHERE AIR BAG BAG DEPLOYED** SHOULD HAVE DEPLOYED BUT DID NOT 1998 All Injuries 335 8 Serious Injury * 115 1 1999 All Injuries 362 20 Serious Injury 131 5 2000 All Injuries 397 24 Serious Injury 143 2 2001 All Injuries 383 17 Serious Injury 118 4 2002 All Injuries 457 24 Serious Injury 173 6 % OF COLLISIONS WHERE AIR BAG SHOULD HAVE DEPLOYED BUT DID NOT 2 0.8 5 4 6 1 4 3 5 3 * Serious Injury is defined on the Abbreviated Injury Scale (AIS) as 3 to 5. An AIS 1 injury is relatively minor. AIS 5 injuries are fatal. ** Air bag deployed as a result of the collision The table shows that, averaged over the 5-year period 1998-2002, the air bags did not deploy when they should have in 4% of the frontal crashes involving all injury levels. The non-deployment rate was 2% for crashes involving serious injury (AIS level 3 to 5). It should be noted that the NASS statistics only document injuries that are apparent at the time of the accident. In many cases the seriousness of an injury may not be recognized for hours or days after an accident. Other serious injuries may not be well represented in the NASS database because * Presenting Author the AIS scale was designed to categorize life-threatening injuries. On this scale the loss of an eye rates as an AIS 2 injury. As “smart” air bag systems are introduced, a rise in the nondeployment figures due to an increase in the sophistication and complexity of the air bag systems may be seen. If the nationwide percentage of vehicles with air bags that have the potential to fail to deploy was 4%, it would suggest that among the 170 million vehicles on the road today, there are millions of defective air bag systems, which will not deploy when needed. This poses a potentially significant risk to the public. Air Bag, Automobile, Non-Deployment C10 A Method for Evaluating the Reliability of EDR Crash Data and Considerations of Consistency Between EDR Crash Data and Post-Accident Artifacts William Rosenbluth, MS*, Automotive Systems Analysis, Inc., 12015 Canter Lane, Reston, VA 20191 By describing these efforts and methodology, the attendee will learn the methods of characterizing crash sensor sensitivity. These methods include an introduction to the identification and analysis of public accelerometer specifications and to public micro controller specifications as can be identified from a teardown analysis of the EDR series to be investigated. This analysis is illustrated with an algebraic derivation of EDR unit data sensitivity using binary and hexadecimal arithmetic. Lastly, the process to derive a cumulative velocity loss from a hexadecimal crash record will be shown for an example. This presentation will impact the forensic community and/or humanity by teaching the attendee to de-mystify the subject of black box data and black box data analysis by seeing practical examples of technical analysis. This will allow the attendees to extend their existing forensic skills, abilities and professionalism by applying them to an area scientific investigation where pre-2000 skills are often inadequate. SYNOPSIS: This analysis illustrates a method of evaluating the reliability crash record data in Event Data Recorder (EDR) and includes a discussion of the consistency of EDR data with the physically reconstructed evidence after a collision. The method described illustrates an engineering analysis to determine the calibration of an electronic crash sensor, and then requires using series of calibrated acceleration pulses impinged on an exemplar electronic crash sensor to document its response to known input pulses. These steps allow the investigator to derive a calibration transfer function for the raw EDR data versus a known calibrated acceleration input. Once the relationship between the saved EDR data and the external calibrated acceleration pulse is known, the subject electronic crash record could be evaluated for consistency with the physically documented reconstruction evidence. LEARNING OBJECTIVES: By describing these efforts and methodology, the attendee will learn the methods of characterizing crash sensor sensitivity. These methods include an introduction to the identification and analysis of public accelerometer specifications and to public micro controller specifications as can be identified from a teardown analysis of the EDR series to be investigated. This analysis is illustrated with an algebraic derivation of EDR unit data sensitivity using binary and 129 hexadecimal arithmetic. Lastly, the process to derive a cumulative velocity loss from a hexadecimal crash record will be shown for an example. THEORY OF THE ANALYSIS: The steps required to accomplish this analysis had to focus on several problems: Retrieving the Crash-Related Data The first problem to be solved was to develop the ability to read the crash-related data inside the appropriate event data recorder (EDR). This required a study of SAE and ISO guidelines and specifications for vehicle networks, and a verification of which particular sub-specification applied in the instant case. This was done using several vehicle network analysis tools as a network traffic monitor for manufacturer/aftermarket scanner equipment doing similar or partial interrogations. In general, the retrieval development process had to include: A. Interrogation of the target EDR so as to obtain the desired electronic information [e.g., EEPROM/Flash-memory hexadecimal data]. This information is often enclosed within interrogation commands and responses, not part of the actual desired information. B. Parsing is the next step. Parsing is the process of extracting the desired electronic information from within the undesired interrogation commands and responses. C. Formatting, the next step is the process of assembling the parsed data into a structured format so that it can be easily read and interpreted. This often involves adding address identifiers and spaces between data elements. D. Translation is the fourth step, is the process of performing an evaluation of the address-identified data elements into meaningful engineering units. Data element translation is usually accomplished in accordance with a standard specification such as SAE J2178-2, SLOT instructions [scaling, limits, offset and transfer function guidelines]. Meaningful engineering units allow the investigator to report such parameter facts as acceleration, timing, seatbelt usage, instantaneous velocity change, cumulative velocity change, etc. E. Interpretation of the translated data is the fifth step. Data interpretation involves evaluation of the reported and translated data with respect to their consistency with complementary investigator findings and conclusions [e.g., reconstruction, bio-mechanics, human factors, etc.]. Deriving a Scaling and Transfer Relationship From the Data. A standard for interpreting ECU data hexadecimal values is given as an SAE SLOT definition for the parameter saved at that address, so that the ECU data can be readily interpreted and used for engineering analysis. In general, information for specific ECU SLOT definitions is not publicly available, so it was required to open the appropriate ECU and photographically document its componentry, specifically, its accelerometer and its micro controller. An example of such a photographic documentation is shown in Figure 2.1. Once the key components are identified, as in this case, industry specifications will reveal the SLOT factors specific to those components. As a tutorial example, A. We assume that the micro controller used an 8-bit A/D converter with a 5,000-volt reference. B. We assume that the accelerometer used has an output sensitivity of 40G/volt, or 1G/0.0250 volt or 0.0250 volt/1G. C. We assume that acceleration A/D counts are represented in EDR memory where each byte (8 bits) is a separate value. For an 8-bit A/D converter as assumed above, it is known that its hexadecimal count range is $00 to $FF (0 dec to 255dec). It can be determined (from manufacturer public specifications) that its full count (255 dec) represents 5.000 volts and that it is a linear radiometric device. Thus, each linear count represents 5.000/255 volts or 0.0196078 volts. For the accelerometer assumed above it can be determined (from manufacturer public specifications) that its nominal sensitivity of that device is 0.0250 volts/G or 1G/0.0250V. 130 Using those two data specifications, the subject EDR data byte acceleration data sensitivity can be calculated as: for each byte of an acceleration record. Some EDRs report their crash-related data in units of direct acceleration per time period (a(t)), some report units of averaged acceleration per time period <a(t)> and others report their data in units of cumulative velocity change per time period time (_ v(t)) . Of course, for any unit time period (tp), the period velocity change, v(t) is no more than the product of a(t) tp, thus, any subsequent validation of EDR Delta V analysis is really validating the analysis of acceleration recording capability. The above discussion assumes that a reasonably complete crash pulse function is being analyzed. However many EDRs may report their data as a different function that incorporates only a portion of, or derivative of, a(t) or v(t). One such value that may be reported is the cumulative velocity at the time of a deploy command, _ v(t)|at fire command. Next the methods and tests to determine and/or confirm such relationships are explored. Conducting Tests to Verify the Above Analysis The ultimate validation of any theoretical analysis is a physical trial of the analysis theory. In this case such a trial consists of impinging a known calibrated acceleration pulse on an EDR and then comparing the EDR record with the known input pulse. For confidence this must be done over several calibration levels around the pulse magnitude of interest. Additionally, if the data acquisition system used to record the impinged acceleration pulse also records an air bag/pre-tensioner squib fire pulse, the time relationship from EDR acceleration record start to squib firing decision can be documented (for the test input acceleration pulse). An example of one of the calibration trial (from a process of several such trials) is illustrated the sequence of Figures 3.1, 3.2, 3.3, 3.4, 3.5. This figure sequence shows the accelerationimpingement fixture, the external recording accelerometer, the external acceleration record, the EDR acceleration record, the superposition of the EDR record on the external acceleration record and the squib fire pulse resulting from that external acceleration input. Lastly, Figure 3.6 shows a superposition of the (calculated) cumulative velocity changes for both the EDR and the external acceleration pulse. That superposition shows that the cumulative velocity changes are essentially congruent (thus proving that the EDR is recording a true representation of the input pulse that it was impinged upon it. Additionally, the time value data for the start of squibfire time shown on Figure 3.5 is also called the time of deploy command. This time can be applied to Figure 3.6 to determine the cumulative velocity change at the time of the deploy command, _ v(t)|at fire command. The above example illustrates a reasonably complete example for a substantially complete whole crash-event. However, a similar analysis could actually illuminate a sub-event. Postulating the sub-event as a whole event would be incorrect. Thus the investigator must consider the consistency and plausibility of the physical reconstruction assessment of the ensemble event with every EDR record analysis. Examples of plausible and implausible EDR analyses are shown to assist the investigator in future such determinations. * Presenting Author C11 Multi-Variable Measurement and Comparison of Rear-Impact Head/Neck Injury Risk for Motor Vehicle Seats Kenneth Saczalski, PhD*, 1440 West Bay Avenue, Newport Beach, CA 92661; Mark C. Pozzi, MS, 8 Paul Road, Cedar Crest, NM 87008; and Todd K. Saczalski, BS, 1505 Verde Valley School Road, Sedona, AZ 86351 Electronic Data Recorders, EDR Analysis, Crash Data Analysis * Presenting Author The goal of this presentation is to present a test protocol, and results, for objective measurement and comparison of motor vehicle seat safety system performance, as it relates to rear-impact head/neck injury risk factors for various size occupants subjected to a high-low range of impact severities (i.e. whiplash range up to 50 kph rear-impact levels). This presentation will impact the forensic community and/or humanity by demonstrating the multi-variable, high-low impact, “side-byside” test protocol, with full vehicle interiors, which provides a more accurate means for comparing vehicle safety system performance as it relates to occupant injury risk measures. Prior studies dealing with evaluation of rear-impact head and neck injury performance of automotive seats have suggested that the more common, but weaker, single-recliner (SR) seat designs (i.e. about 3.2 kN strong), which tend to collapse rearward during rear impact, provide improved occupant protection over the much stronger and available “beltintegrated” seat (BIS) designs (i.e. about 14.5 kN strong), for impact severities ranging from low velocity “whiplash” levels (i.e. 17 kph or less) on up to more severe rear impacts of 40 to 50 kph. It has also been suggested in these studies that out-of-position (OOP) occupants in the stronger BIS designs are at greater risk of injury than if they were seated in the weaker SR seat designs. Unfortunately, these studies did not test the weaker seat performance within the full constraints of the vehicle interior and rear occupant space. These constraints include, among other things, non-yielding rear seatbacks, limited rear occupant space, hard rear surface structures like package shelves, and rear occupants themselves (such as children and infants). Also, other factors that are not likely to affect the BIS design, but can further degrade the performance of the collapsing SR seats (such as intrusion of rear seatbacks, vehicle pitch/yaw motions, and offset rear impact) have not been examined in these earlier studies. This current study evaluates head and neck injury performance of each seat design by using a complete vehicle interior, with “side-by-side” testing of both the typical collapsing SR and strong BIS designs, for 3 sizes of restrained surrogates subjected to a “high-low” range of rear impact severity. Neck injury performance is based on the “percent risk of AIS (Abbreviated Injury Scale) 3+ injury” derived from the NHTSA (National Highway Traffic Safety Administration) combined load “Nij” values, calculated from the measured surrogate response. Head injury risk is based upon the HIC (Head Injury Criteria) curve for “percent population at risk of AIS 4+” head injury. A typical 4-door family sedan vehicle, with full interior, was used as the baseline vehicle. Both “sled-body buck” tests and 2 complete “vehicleto-offset barrier” pole impact tests were used. A total of 3 sled-buck tests were run at a low “whiplash” severity level (17 kph) and 6 were run at the severe high impact velocity change (50 kph) (repeat tests were run at the high impact levels). The crash pulse applied to the body-buck system was matched to the crash pulse of the actual vehicle. Three occupant sizes (i.e. 5th Female (50 kg), a 50%tile Male (80 kg), & a large Male (50%tile Male ballasted to 110 kg)) were tested for each of the 2 velocity change levels. Each surrogate was instrumented with head, upper neck and chest instrumentation. Lap belt loads were also measured. In all cases the surrogates were leaned forward “out-of-position” (OOP) from the headrests with a gap of 5 to 6 inches, to examine effects of occupants in non-optimum seating positions for both seat types (SR and BIS). The effects of “rear seatback intrusion” and “vehicle pitch” that could adversely effect the weaker SR seat were excluded in the “sled-buck” tests, so that it would be possible to evaluate the collapsing SR seat performance under optimum conditions. Ultimately, the adverse effects of rear seatback intrusion, etc, were 131 evaluated with the actual “vehicle-to-barrier” tests. These tests were run at the high severity level, with the heavy surrogate in the weaker SR driver seat in one test, and the other test run with the heavy surrogate in the BIS version of the driver seat. In some of the sled-buck tests a 6-year-old surrogate was placed in the rear bench seat to study other hazards of the collapsing SR seat design. The data and results of the “side-by-side” seat tests are summarized in several tables. Included in each table is a category for “% Risk of AIS 3+ Neck Injury Potential,” and “% Population at Risk of AIS 4+ Head Injury.” What the results indicate is that for the strong BIS design there is no significant risk of head, neck or chest injury at either the “whiplash” 17-kph levels or the more severe 50-kph range. On the other hand, the weak SR seat demonstrates “serious and dangerous” risk of head and neck injury in the “severe” 50-kph ranges, for all size occupants, even with optimum headrest height and no adverse rear seatback intrusion or vehicle pitch/yaw effects. Also, common vehicle mounted belts were ineffective in restraining all size occupants in the collapsing SR seat design at high severities. Finally, the effects of rear seatback intrusion, and vehicle pitch/yaw, as well as impact offset, clearly increases the injury risk factors for the SR seat, but have virtually no deleterious effect on the BIS designs. Rear-Impact, Motor Vehicle Seats, Head/Neck Injury Risk C12 A Semi-Trailer/Ambulance Collision on a Dark and Icy Night: Proof of a Bizarre Bounce Contradicts Witness Statements and a Police Report Joseph A. Keierleber, MFA, MTC Forensics, 54 Pleasant Avenue, Peaks Island, ME 04108; and, Thomas L. Bohan, PhD, JD*, MTC Forensics, 54 Pleasant Avenue, Peaks Island, ME 04108 The goal of this presentation is to describe an unusual motor vehicle crash in which misleading eyewitness statements and a careless police report nearly obscured the true cause of the collision. In addition to being an object lesson in the unreliability of eyewitnesses, this case should also serve to instruct attendees on how conclusive statements, even those of official police reports, may be the result of prejudice and uncritical thinking. Finally, it will reemphasize the rewards of iterative investigation techniques. This presentation will impact the forensic community and/or humanity by demonstrating the importance of continued analysis followed by reexamination of physical evidence suggested by that analysis is an essential part of the forensic process and can yield big returns. The Crash: A northbound ambulance on an emergency run with its blue lights activated collided at highway speed with the side of a southbound semi-trailer hauling 45,000 pounds of paper. The ambulance came to rest in the middle of the highway, rotated about 90 ccw and the tractortrailer came to rest alongside the snow bank lining the southbound side of the road, with the rear of the trailer several feet into the snow bank. Road Conditions: The highway sloped downward from north to south. The temperature was near freezing, with a drizzle falling and freezing along the portion of the road to the north of the crash site, but not to the south, from where the ambulance was coming. Driver Statements: (1) The ambulance driver stated that as the ambulance approached the tractor-trailer the latter jackknifed, its trailer swinging around to block the northbound side of the road all the way to the guardrail. (2) The truck driver said that he had slowed and pulled to the right when he saw the ambulance approaching and just after the ambulance had passed him he heard a crash and, looking back, saw that the ambulance had run into the side of his trailer, which was tracking directly behind the tractor. Eyewitness Statements: All four persons from abutting residences who came forward were insistent that it was the fault of the ambulance driver. The witnesses depicted the ambulance as traveling at a high rate of 132 speed and veering into the truck, which they said was traveling slowly and entirely in its own lane. A northbound driver said that the ambulance lost control after passing a vehicle at a high rate of speed, after which it crashed into the trailer, which was in its own lane of travel. Another driver, who disappeared right after the crash, said that the tractor-trailer had jackknifed, causing the crash. Police Report: The crash occurred in early evening, after dark. The local police investigator concluded that the ambulance had caused the crash and so stated to the region’s newspaper, which ran a story to this effect, amplifying it with accounts of the repeated complaints reportedly made by residents concerning ambulances speeding past their houses. It also included the erroneous police statement that the speeding ambulance was just returning to its home station and not on an emergency run. Physical Evidence: The most striking physical evidence was a 50-foot stretch of chewed-up guardrail on the northbound road edge was chewed-up. The top of the guardrail was originally a few inches higher than the bottom of the anti-underside bumper (ICC bumper) on the trailer. That bumper showed massive damage to all areas and it was clear that it had been struck either by the ambulance or by the trailer tires, the carriage for which had been dislodged from its mounts on the trailer’s underside. The damage to the trailer, including paint transfer, showed that the ambulance had struck it approximately in the middle and then had slid along it until coming into contact with the trailer’s front left outboard tire. (The trailer had dual tandem axles positioned near its aft end.) The ambulance displayed massive damage to the front left quarter extending back to the region behind the driver’s seat. There was essentially no damage to the right side of the ambulance. The Problem: Although the guardrail damage upstream from where the vehicles came to rest strongly suggested that the truck had indeed jackknifed, the adamant statements of the witnesses initially cast doubt on that point. One of the witnesses even asserted that the ambulance post-impact caused the chewed up guardrail. However, a careful examination and reexamination of the ambulance turned up no portions of it that could have ridden the rail in the fashion required for that damage. The police report did not take that damage into account at all; by his own admission, the investigating officer never examined it closely. He was willing to say that both vehicles were slightly over the centerline and that the responsibility was therefore evenly divided. The biggest sticking point to explaining the crash as resulting from the jackknifing that blocked the ambulance’s path was the lack of damage on the right side of the ambulance. If the crash had occurred with the trailer’s aft portion riding the guardrail, it would seem that the ambulance would have been forced into the guardrail; yet there was no marking on the ambulance to suggest that it had hit the rail at all. Also, it was not clear that the impact of the ambulance against the trailer would have provided the momentum necessary to swing the trailer back off the road on the other side. The Solution: The road was so slippery at the time of the crash that the vehicle dynamics were akin to air table physics. If the truck had been jack knifing so as to bring the ICC bumper into contact with and past the guardrail on the opposite side of the road, the swinging of the trailer would not have stopped at that point, but would have continued so as to bring the outer sidewall of the trailer’s left rear-most tire into contact with the guardrail. When that tire was re-examined, two horizontal wear lines at the height corresponding to the heights of the two guardrail protrusions were found. The portions of the rim corresponding to the respective centers of the two wear marks were scored in a way consistent with them having rubbed against the steel guardrail. The fact that there were two discrete wear bands had been overlooked before because of paint in the same area of the tire. Also, a faint black discoloration of the guardrail was found toward the south end of the chewed-up section. As it turned out, the impact between the front left outboard tire and the ambulance had forced the two left-side axles together so that none of the trailer tires could rotate following the impact. It was for this reason that the two parallel scuffmarks remained horizontal to the ground. This additional evidence of contact between the trailer and the guardrail made it clear that when the truck driver slowed and pulled to the * Presenting Author right because of the approaching ambulance the trailer tires locked up on the very slippery surface, resulting in the jack knife. The jack knife went unnoticed by the truck driver, who was intent on the approaching ambulance. Meanwhile, the jack knife continued, the trailer tires not rotating, until the rear end of the trailer hit the guardrail on the opposite side, ultimately bringing its rear, left tire into contact with the rail. The swing continued, causing the tire to be compressed, until the rim came into contact with the rail, halting further swinging. When the swinging ceased, the trailer moved outward from the rail as the result of the force of the compressed tire on the rail. Making reasonable geometric assumptions about the compression, as well as the physical assumption that that part of the tire away from the rail did not change in shape, the reduction in volume of the tire could be estimated. The kinetic energy given to the trailer by its tirerelated rebound from the rail was then equated to the tire pressure (which would have remained essentially unchanged because of the small relative volume change) multiplied by the change in volume as the tire pushed outward W = p ∆V. This quantity turned out to be approximately 1,200 ft-lb, which in light of the trailer weight and moment of inertia about the fifth wheel, indicates that during the rebound the end of the trailer initially moved away from the guardrail at a speed of about 10 feet/second. As a result, an opening appeared between the trailer and guardrail by the instant of impact. In addition, the trailer was swinging away from the ambulance’s lane at the time, and as a result of these two factors, the collision did not force the ambulance into the guardrail. Furthermore, this explains why, when the truck driver heard a bang after passing the ambulance, he looked back to see his trailer tracking behind the tractor. Finally, this explains how the trailer fetched up in the snow bank along its own side of the road, which was part of the basis for the witnesses and police investigator being convinced that the truck could not have been at fault. Accident Reconstruction, Jackknife, Eyewitness C13 Forensic Testing and the Characteristics of Seat Belt Webbing Force Limiting Expansion Loops Kurt D. Weiss, MSME*, Automotive Safety Research, Inc., 5350 Hollister Avenue, Suite D, Santa Barbara, CA 93111-2326 After attending this presentation, attendees will have an understanding of seat belt force limiting expansion loops, the associated testing methodologies, and the force-displacement characteristics of several designs incorporated in late model vehicles. This presentation will impact the forensic community and/or humanity by demonstrating expansion loop characteristics and providing several data points useful in analyzing vehicle occupant interaction with the seat belt system. THEORY OF THE ANALYSIS: Seat belts remain the primary occupant restraint system in passenger vehicles. In the event of a collision, 3-point lap and shoulder belts help to reduce the risk of injury due to impacts with interior components or ejection. Seat belts offer ride-down by providing restraining forces early in the collision event, thereby decreasing the relative velocity between the vehicle and occupant. Seat belts also help distribute restraining forces over the skeletal areas of the body. In an effort to modify occupant kinematics and to reduce the injury producing restraining forces, seat belt designs often incorporate expansion loops. An expansion loop is a commonly used force-limiting device whereby the seat belt webbing is folded along the short axis, and stitched in place by parallel rows of threading. The thread material and stitch pattern used to make the rows of stitching will tear at predetermined webbing tension levels. When the stitches tear, the webbing fold will open * Presenting Author thereby lowering the tension in the webbing. A warning label is often visible after the stitches tear. Replacement of the seat belt assembly in this condition is strongly advised. Testing was conducted on new and used front outboard seat belt assemblies of three different vehicle models to evaluate the percent elongation of the webbing, and to determine the force-displacement characteristics of the expansions loops. The instrumentation and scientific methodology used were typical and standard for this type of forensic testing and data acquisition. SEAT BELT WEBBING TEST SET-UP: 14 type 2 seat belt assemblies were obtained for testing. Six new assemblies were purchased from local dealers, and eight used assemblies were obtained from salvage vehicles. Preparation of the samples included separating the webbing from the retractor mechanism. The rubber-like escutcheons that often encase the expansion loops were preserved and left intact for use in the second of two test series. TEST SERIES: Two series of tension tests were conducted. The first series (Series A) contained 14 tests, one test per webbing sample. The test protocol was designed to determine the percent elongation of the webbing, and did not include the effect of the expansion loop. Webbing samples tested were sectioned from the portion of webbing nearest the seat belt retractor. Webbing ends were held by split drum grips of the type specified under Federal Motor Vehicle Safety Standard (FMVSS) 209. One split drum grip was rigidly anchored to the test bench, while the other grip was secured to the crosshead of the test machine. Tension in the webbing was applied by raising the crosshead. A webbing pre-load of approximately 222.4N was used. The grip separation rate was 51mm per minute. The force was measured by a load cell. Elongation of the webbing was determined with a laser extensometer, as well as with an engineering scale, when the tension in the webbing rose to 11,120N. The second test series (Series B) contained 14 tests, one test per webbing sample. These tests were performed to evaluate the forceextension characteristics of the expansion loop of the lap belt. The webbing section tested in this series was the portion nearest the outboard lap belt anchor. One end of the sample was secured to the test bench using the existing anchor bracket to which the webbing was sewn. The expansion loop remained concealed in its rubber-like escutcheon to maintain the original in-vehicle condition. The other end of the webbing sample was held by a split drum grip attached to the test machine crosshead. The grip separation rate was 100mm per minute, a rate commonly used by the seat belt manufacturing industry. Force was measured with a load cell. The peak force to tear each row of stitching was determined. As tension in the webbing sample increased, the rows of stitching in the expansion loop subsequently failed. As the rows of stitching were torn, tension in the webbing decreased as a small amount of webbing was added to the sample overall length. This phenomenon produced a saw tooth-like force-displacement curve. Sample testing was stopped after all rows of stitches were completely torn. TEST RESULTS: The results of Series A varied by vehicle model. For new samples, the webbing elongation was between 5.2% and 14.8% using the laser extensometer, and between 5.9% and 13.6% using the engineering scale. For used samples, the webbing elongation was between 5.5% and 11.4% using the laser extensometer, and between 5.8% and 10.7% using the engineering scale. The results of Series B were dependent on the expansion loop design, i.e. by the fold pattern, the number of rows of stitching, the thread material, and the stitching pattern. One expansion loop design added an average 218mm of webbing at an average peak force of 2733N for new samples tested. The used samples added an average 267mm at an average peak force of 1900N. Another expansion loop design added an average 238mm of webbing for the used samples. The first three rows of stitching yielded at an average peak force of 5067N and the remaining rows yielded at an average peak force of 3244N. The testing of the new sample in this vehicle model 133 resulted in a failure of the webbing during the test. The results of this sample were suspect and therefore not included. The last expansion loop design added an average 248mm of webbing for the new samples. The first three rows of stitching yielded at an average peak force of 5910N and the remaining rows yielded at an average peak force of 4225N. For the used samples, an average of 231mm of webbing was added. The first three rows of stitching yielded at an average force of 4943N, and the remaining rows yielded at an average peak force of 3600N. CONCLUSION: Expansion loops are often incorporated in seat belt designs to modify occupant kinematics and reduce occupant-restraining forces during the collision event. Expansion loops vary among vehicle models and each design exhibits unique force-displacement characteristics. Forensic testing of seat belt expansion loops can be used to quantify the force necessary to break the stitching and measure the amount of additional webbing added to the seat belt assembly. Subsequent testing results will be offered in future publications. Seat Belt Webbing, Force Limiters, Expansion Loops C14 Truck Unloading Caused Fatal Bunker Retaining Wall Tipover Thomas L. Boyer, PE*, and John A. Talbott, BS, PE, Talbott Associates, Inc., 7 Southeast 97th Avenue, Portland, OR 97216-2498 The goal of this presentation is to illustrate the investigations, measurements, testing and analysis, which determined the cause of a bunker wall overturning and resulted in a large settlement for the decedent’s estate. This presentation will impact the forensic community and/or humanity by demonstrating a methodology for analysis of unusual interactions of vehicles and structures, which may usefully be applied to other events of similar character. A company in Molalla, Oregon, specialized in preparing and selling bagged bark dust and other soil amenities. The bulk bark dust was received from a single supplier, who delivered the bark dust in tractor-trailers. The bark dust was stored in open-top bunkers constructed of concrete blocks 2’ x 2’ x 6’ with an interlocking tongue-and-groove which were stacked on the asphaltic concrete to provide the sidewall enclosure into which the trucks unloaded. The semitrailers had “walking floor” live bottoms for self-unloading. One of those bunkers was fairly close to a metal building on the property, and because there were no other sanitary facilities close by, there was a “Port-a-Potty” situated between the retaining wall of the bunker and the metal building. The standard procedure was for the truck driver to back his truck up so that the trailer was about 6’ from the wall of the bunker and then for him to wait for an observer from the processor’s crew to stand by and watch the unloading and then to release the tailgate and release the brakes and begin the unloading process. The live bottom of the semitrailer would slowly move the bark dust cargo rearward, forcing the top-hinged tailgate open, and depositing the chips on the asphaltic paving in the bunker. Then, as the expelled cargo mounded behind the trailer, the tractor and semitrailer would be forced forward, allowing the mound to grow forward. At completion of unloading, the tractor-trailer would be driven forward to allow the tailgate to clear the mound and close. The paper demonstrates a methodology for analysis of unusual interactions of vehicles and structures, which may usefully be applied to other events of similar character. On the occasion of the accident, a woman was in the Port-a-Potty; an observer from the processor’s crew was not immediately available, and the truck driver decided to proceed with unloading anyway. As the unloading proceeded, the wall of the bunker tipped outward, partially crushing the Port-a-Potty against the building and exerting such pressure on the woman who was then standing that she was asphyxiated by compression of her chest and abdomen. The question was why the wall tipped. 134 Talbott Associates visited the site, measured the blocks of which the wall was formed, and measured the slope of the asphalt surface over which the tractor would have passed during its unloading. There was observed no foundation or soil failure. Next, the truck was examined at the supplier’s yard, loaded in the manner and to the extent of the subject case. The slope of the drive on which the truck was situated was measured along its full length, and then with its brakes released, it was pulled by another truck while a dynamometer measured the force required to cause its motion. The trailer’s top pivoting tailgate was measured, as were the general tractortrailer dimensions. Observation was made of concrete abrasion marks on parts of the trailer gate. Samples of the different grades of bark dust were taken and were used to determine its density and coefficient of internal friction, shear strength, and its lateral pressure coefficient. Because the bunker wall had been moved and replaced since the accident, no artifacts of the accident were discoverable on the wall itself. Structural analysis of the stability of the concrete wall revealed that it had a huge factor of safety against overturning for retaining the bark dust and wood chips. Further analysis revealed that if the tailgate did not contact the wall, the force that could have been exerted through the chips could not have been enough to tip over the wall. Analysis also indicated that even if the tailgate had made contact with the wall, the geometry and mechanics of the unloading bark would provide a forward horizontal force on the trailer which exceeds the rearward horizontal force at the tailgate to wall contact. If the brakes of the tractor and trailer were released, the force required to move the trailer forward and thus the force applied to the wall was found to be insufficient to cause a tipover of the wall even with the combined effect of the lateral pressure of the chips against the wall. Therefore, the conclusions were that the trailer was stopped too close to the wall, the brakes were still applied, and the unloading proceeded without the driver watching the process and without the observer, which was required by the normal procedure. A videotape of an unloading procedure taken by the Oregon Occupational Safety & Health Administration was reviewed, and computerized animations were performed to illustrate the process of the accident. Suit was filed on behalf of the estate of the decedent woman, and after the defendants’ counsel’s viewing the animation and other evidence; a substantial offer in settlement was proffered and accepted. Unloading, Wall, Tipover C15 Determination of Driver Identity in a High-Speed Vehicle Rollover: Vehicle Occupant Injury Patterns and Vaulting Velocities Correlated With Safety Defects and Accident Reconstruction Mark C. Pozzi, MS*, Sandia Safety Sciences, 8 Paul Road, Cedar Crest, NM 87008; and Todd K. Saczalski, BS*, TKS Consulting, 1505 Verde Valley School Road, Sedona, AZ 86351 After attending this presentation, attendees will understand forensic science techniques for determining vehicle occupant trauma pattern and vehicle collision damage, and correlation of this with scene evidence, vaulting velocities and vehicle dynamics, to assist in determination of driver identity. Attendees will also understand the application to general biomechanics, accident reconstruction, vehicle safety investigation, static and dynamic test techniques and evaluation. This presentation will impact the forensic community and/or humanity by demonstrating a new method of utilizing static and dynamic testing as well as previously conducted non-case-related tests to determine critical factors in accident reconstruction, especially for determination of driver identity. Determination of driver identity is a well-recognized * Presenting Author demand for law enforcement, prosecutors, insurance companies, accident investigators, and medical examiners. A wide variety of forensic evidence identification and analysis techniques were utilized in this investigation, which occurred several years after the event. LEARNING OBJECTIVES: The authors will present reliable forensic science techniques in analyzing trace evidence from photos, medical records and physical examination of a vehicle and accident scene. Via the scientific method, demonstrate vehicle safety failures, as well as how to utilize available test information to determine performance of vehicle subsystems and components in a unique collision. PROPOSITION: Via the scientific method, show correlation between specific vehicle damage, occupant vaulting energy, and occupant injury, at specific points during a high-speed rollover event, as a reliable means to determine vehicle occupant identity. SYNOPSIS: A forensic case study is evaluated which involves a single-vehicle high-speed fatal rollover accident with both occupants ejected. It was initially unknown which occupant was driving the vehicle, despite investigation by the state police and the regional medical examiner. The surviving occupant was severely brain injured with no memory of the accident. There was no witness to the rollover, which involved extensive off-road vehicle travel with significant vertical increase in elevation, followed by falling down a rocky cliff. No fingerprints, hair, blood, skin smears, fibers, fabric transfer, or other organic samples were positively identified. Mud deposits on the vehicle interior and the lack thereof on one occupant’s shoes, as well as specific evidence (or lack thereof) involving steering column, dash, control pedals, windshield, seats, seat belts, door interiors, side and rear windows, and headliner were analyzed and identified by the authors on the vehicle interior. Evidence (or lack thereof) on door latches, door structures, door window openings, exterior sheet metal, exterior mirrors and trim, axles, and numerous other exterior vehicle structures was analyzed and identified by the authors. The occupants were ejected from different vehicle portals, at different locations, as a synergistic result of restraint system failure, seat failure, and door latch failure, correlated with vehicle collision and rollover dynamics. Occupant trauma pattern and severity was determined from photographs and review of autopsy and medical records. Occupant trauma pattern/severity and vehicle dynamics were accurately matched to vehicle interior and exterior evidence, occupant vaulting velocities, as well as occupant and vehicle final rest positions. The foregoing was correlated to evidence deposited by the vehicle, as well as each of the vehicle safety failures, to determine which occupant was ejected from which portal, at a specific place and time. Static and dynamic testing by the authors and others was correlated with dynamic vehicle collision and rollover mechanics to also prove occupant ejection dynamics, occupant injury or lack thereof, and occupant location within the vehicle. This involved use of: A) Static and dynamic seat tests to determine that a specific seat was unoccupied at a certain point in the rollover event. B) Dynamic crash and sled tests to prove restrained and unrestrained occupant kinematics, injury level and restraint system failure. C) Static seat belt tests with human surrogates to determine likelihood of restraint system use by a specific individual. The vehicle rollover path was accurately matched with location and type of each significant collision/ground contact, vehicle exterior damage, and vehicle parts/debris deposits. Each significant collision/ground contact was then correlated with vehicle velocity and safety failures as well as occupant ejections, vault velocities and trauma patterns. SUMMARY/CONCLUSION: Reliable, positive identification of the driver was made via multiple cross-correlation of all the foregoing evidence with biomechanical and vehicle dynamics analysis. What may be a new technique of correlating static and dynamic test data to a specific real-world crash event was developed. Biomechanics, Reconstruction, Rollover * Presenting Author C16 Conspicuity vs. Visibility in Accident Reconstruction James B. Hyzer, PhD*, Hyzer Research, 1 Parker Place, Suite 330, Janesville, WI 53545-4077 The goal of this presentation is to show that the conspicuity of a hazard is a critical factor in reconstructing the visibility aspects of a collision between a vehicle and a hazard. This presentation will impact the forensic community and/or humanity by demonstrating a better understanding of conspicuity and visibility in reconstructing the visibility aspects of vehicle/hazard collisions. The objective of this paper is to discuss and analyze the factors that affect a driver’s ability to visually perceive and then recognize hazards such as pedestrians, objects, and obstructions ahead under varying conditions of background clutter. The outcome will be to show that the conspicuity of a hazard is a critical factor in reconstructing the visibility aspects of a collision between the vehicle and the hazard. A common cause of roadway accidents is the driver not seeing a hazard in time to respond and avoid collision. It is not uncommon for drivers who have hit pedestrians, for example, to report that they “heard the thump but never saw him” or that they “didn’t see him until it was too late.” When presented in court, the outcome of the case may depend on whether the jury believes the driver’s testimony or concludes that since the pedestrian was in plain sight, the driver must be lying. The job of the expert, then, is to determine whether the signal value of the struck pedestrian was sufficient relative to the driver’s visual field to assuredly capture the attention of all reasonable alert drivers exercising ordinary care with respect to lookout in time to enable them to respond in time to avoid collision. Through a review of published scientific literature, it will be shown that it does not follow that simply because a pedestrian, obstacle or obstruction is in plain sight that it will necessarily be perceived as a hazard and responded to in time by all reasonable alert drivers. Under low-light-level and/or nighttime conditions, and at locations with no or minimal background clutter or visual confusion (e.g., rural settings), the visual perception and recognition of a hazard in the roadway simply requires that it be either more or less luminous than its immediate background and have a sufficient luminance and contrast to be distinguishable from its background. Contrast is related to the difference in the luminance of an object of interest and the luminance of its immediate background. Contrast sensitivity is quantitatively equal to the reciprocal of contrast threshold and represents a measure of an observer’s ability to discriminate different levels of contrast. The contrast sensitivity of the human visual system decreases with age and with lower light levels. Other factors that need to be considered in determining the visibility of a hazard under low-light-level and/or nighttime conditions are such things as observer expectancy, the age of the observer, exposure time, disability glare, light adaptation, purkinje effect, positive vs. negative contrast, headlight beam-pattern, and the relative geometry of the driver and headlights with respect to the hazard. Under daytime and nighttime conditions at locations with significant background clutter and/or visual confusion, hazards can be perceived and recognized when they differ from their surroundings in such aspects as size, shape, luminance, color, motion, texture or some other visual disparity. Conversely, hazards that are visually identical to their surroundings cannot be seen or recognized and are said to be perfectly camouflaged. To correctly express these concepts to a jury, it is important to understand the relevant terminology as it relates to visibility. To detect an object means to discover or determine its presence. A detected object that is also a hazard, however, may not necessarily be recognized as a hazard. To see the object, simply means to perceive it by the eye or by vision. To perceive it means to become aware of its presence through the senses (here by vision). An object is visible if it is capable of being seen. An object is conspicuous if it attracts or tends to attract the attention of an observer so as to be readily discovered by vision. Conversely, an object is inconspicuous if 135 it is not readily noticeable or discoverable by vision. The term conspicuity, then, is the capacity of an object to stand out in relation to its background so as to be readily discovered by vision Even though a hazard may be in plain sight and visible, it must be conspicuous relative to its surroundings to be seen in sufficient time by all drivers. Hazards that are more conspicuous are going to be perceived quicker and therefore at greater distances than hazards that are less conspicuous. At the extremes, hazards that are highly conspicuous should be seen at the greatest possible distances and hazards that are perfectly camouflaged will not be seen or recognized at all. In conclusion, since it is clear that hazards such as pedestrians, objects and obstructions that are more conspicuous can be seen earlier and therefore at greater distances than hazards that are less conspicuous, and since it can be shown that inconspicuous or perfectly camouflaged hazards may not be seen at all, then it follows that hazards that are less conspicuous can in fact be shown to be the primary contributing cause of being hit by vehicles with drivers exercising ordinary care with respect to lookout. Conspicuity, Visibility, Accident Reconstruction C17 Facial Comparison of Persons Using Non-Standardized Image Material Arnout C. Ruifrok, Ivo Alberink, PhD, Mirelle I Goos, MSc, and Jurrien Bijhold, PhD, Netherlands Forensic Institute, Volmerlaan 17, Rijswijk, 2288 GD, Netherlands The goal of this presentation is to describe quality issues and a standardized procedure to perform facial comparisons, in order to make the process of performing facial comparison as objective and consistent as possible. This presentation will impact the forensic community and/or humanity by presenting quality issues and methods for visual comparison. Facial recognition and comparison is still one of the ‘promising’ biometric systems considered, and even implemented, in security systems. Error rates at settings with equal percentage false accepts and false rejects (equal error rates, EER) of 5-10% are considered reasonably good, especially with long time-lapse (a year or more) between enrollment and surveillance. Four aspects of image acquisition are of major importance for automated facial recognition: background, lighting, camera position, and facial expression. The more variability in one of these factors, the higher the error rates. Most images of surveillance cameras are taken under suboptimal conditions at best: backgrounds vary, lighting is poor, camera position is mostly with a downward angle, facial expressions are variable, and added to that people wear hats or disguises, and finally most images are saved at high compression levels. It is clear from the above, that biometric systems for facial recognition are not appropriate for forensic applications. Although maybe useful in the investigational stage, the error rates, even in reasonably well-standardized images, are too high for final legal proof. This means that visual matching does the final confirmatory check of the identity of a person. Especially in criminal cases, where the available evidence mostly is limited, but the correct identification of the criminal is crucial, the final decision will also be made by means of a visual check. However, this process is still subjective, and clear guidelines on how to perform such a comparison are hard to find. Therefore, the authors have developed procedures to standardize facial comparisons as much as possible depending on the material available. Preferably, a comparison will take place with pictures taken from the same camera position, and from the same distance. If a person and original camera equipment are available for comparison pictures, a three-point matching method is used to position the person according to the available pictures, and a more robust comparison can be made. This also provides the opportunity to validate the comparison using the actual system: also 136 5-6 foils are imaged under the same circumstances, and analysis results of the foils and the suspect are compared. This will give an indication of the reliability of the facial comparison with the available system set-up. If the original equipment is not available, or the position of the person is hard to estimate, a 3-D laser scanner can be used to build a 3-D model, followed by calculation of the most likely camera position and properties. This provides the opportunity to position suspect and perpetrator images alike, improving the comparison process. However, in many of criminal cases, no additional comparison pictures can be made (e.g., when a crime suspect is still at large), and comparisons have to be performed using pictures from different time periods, camera positions and camera distances. Therefore, the authors have developed a procedure to standardize facial comparisons as much as possible. The method comprises description of general information concerning the material, and a step-by-step comparison and scoring of general facial features (contours, relative measures, and positions), specific features (eyes, nose, ears, mouth, neck and throat), facial lines, folds and wrinkles, and typica like scars, moles, tattoos, and piercings. This approach has been compared the approach used in other countries. From this study it is clear that although the general approach is similar, the level of detail and the reporting of results can be quite different in different countries. In general it is recognized that direct measurements are not suitable for facial comparison, that consistent judgment of images is hard to achieve, and that there is a lack of statistical data for facial features. Implications and possible solutions to some of these issues are presented. Facial Comparison, Identification, Objectivity C18 Body Length Estimation From Surveillance Video: Procedures and Validation Bart Hoogeboom, MS, Ivo Alberink, PhD, Mirelle I. Goos, MS, Derk Vrijdag, MS, and Jurrien Bijhold, PhD, Netherlands Forensic Institute, Volmerlaan 17, Rijswijk, 2288 GD, Netherlands The goal of this presentation is to describe procedures and validation techniques for estimation of the height of a person in surveillance video. The Netherlands Forensic Institute uses a technique with 3-D computer models to estimate the length of a person in a surveillance video. The 3-D-computer model is made with commercial photogrammetry software from photos taken at the scene. The model is then imported into commercial software for 3-D modeling and animating. The model can be viewed from every desired perspective in this software. This implies that the model can also be viewed from the same perspective as the camera that has taped the (video) image. To find this perspective, the software is provided with a camera match algorithm. This algorithm calculates the position and field of view of the camera through the similarity of points between the image and the 3-D model. The user in both the image and the 3-D model must point out these points. The model can now be looked at through a virtual camera with the video as a background. By positioning a 3-D modeled object in the 3-D space in such a way that they fit the person in the image, the height of the person is measured. Mistakes and inaccuracies in this procedure are caused by: • The quality of the 3-D model • The choice of the virtual camera • Uncertainties in the interpretation of the (video) image. (Which pixel on which video line belongs to a certain point of the 3-D model?) Inaccuracies in the 3-D model will systematically affect all the measurements. A mistake or inaccuracy in the interpretation of the (video) image can be random, but also a systematical error from the investigator. For this reason it is wanted that the camera match and the measurement is repeated several times by several investigators. * Presenting Author Reference images are made to validate the measurements and to get a better understanding of the measuring errors. This means shooting images with the same equipment and at the same location that was used for the questioned images. The shooting takes place under controlled circumstances and the camera view is checked for changes. If the camera view has been changed compared to the questioned image, the view will be restored as good as possible. Reference objects with a known size and location are placed in view of the camera. Also, several persons will stand still at known locations. At the lab of the Netherlands Forensic Institute, these reference images are treated the same way as the questioned images. When performing a height measurement on a person in a (video) image it is necessary that the person be fully displayed. In other words: both the head and the feet of the person should be visible in the image. The gait and pose of a person is of great influence to the result of the height measurement. For this reason the aim is to use an image in which the person stands tall and still. When this is not the case an extra error should be taken into account for the height measurement. The quantity of the error for not standing tall can only be estimated for every individual case. However, the influence of the gait of a person to the height measurement has been studied by several people [1,2]. These studies show a maximum variation of the measured height of a person of 8 cm. At this moment there is a study going at the Netherlands Forensic Institute to the influence of the pose of a person, camera view, lighting, and also interpretation of the images by the investigator on the result of height estimation. One of the experiments includes measurements on about twenty people shot by the surveillance cameras from the institute. On the day they were shot on camera and where they were also measured in real live by the investigators. The purpose of this study is to narrow the errors in future cases on height measurements. Literature: [1] David Compton, Clair Prance, Mark Shears and Christophe Champod, A Systematic Approach to Height Interpretation from Images, in Proceedings of SPIE Vol. 4232, Enabling Technologies for Law Enforcement and Security, 2001 [2] Antonio Criminisi and Andrew Zisseman, Luc van Gool, Simon Bramble and David Compton, A new approach to Obtain Height Measurements from Video, in Proceedings of SPIE Vol. 3576, Investigation and Forensic Science Technologies, 1998 Height Measurement, 3-D Computer Models, Surveillance Video C19 Questions About the Integrity and Authenticity of Digital Images: A Review of Case Reports From the Netherlands Forensic Institute Zeno Geradts, PhD*, Derk Vrijdag, BSc, Ivo Alberink PhD, Mirrelle I. Goos, MSc, and Arnout Ruifrok, PhD, Netherlands Forensic Institute, Volmerlaan 17, Rijswijk, 2288 GD, Netherlands This presentation will impact the forensic community and/or humanity by developing standard operating procedure for determining the authenticity and integrity of a digital image file. For forensic casework the question of authenticity has to be answered if a certain image has allegedly been made with a specific digital camera. Another question that may be asked is if two images have been made with the same camera. In order to answer this question pixel defects, and information from the headers and footers of image files can be used. Furthermore, the method of examination of pixel defects combined with headers and footers is useful for integrity research: finding traces of manipulation (e.g. cut and paste) of the images. A digital image is composed from a matrix of pixels (picture elements). For capturing a digital image a Charge Coupled Device (CCD) is * Presenting Author used in a camera. When manufacturing large CCD arrays, they sometimes contain defects. A defect is visible in the image as a pixel defect if the CCD element has a different light sensitivity compared to the surrounding CCD elements. For the examination of pixel defects the authors have developed a standard operating procedure in forensic casework. The examination has two approaches. If the camera is available, test images will be made with the camera with a white, gray, or a black surface. These images are used as a reference set. If the camera is not available, one set of images is used as reference set. In some casework the pixel defects could be visualized without averaging or image processing, since they were visible in the images themselves without any processing. However, for visualizing the pixel defects it is often necessary to add and average the intensities of the images. As a result of this, fluctuations in the images due to the image itself will be averaged. In order to visualize the pixel defects a filter, for instance a median filter can be used. The locations of the pixel defects in the reference images are compared with the location of the pixel defects of the questioned images. If the locations of the pixel defects agree with each other, this provides strong support for the hypothesis that they have been made with the same camera. The conclusions are not quantitative however, since not enough statistical data is available from the randomness of pixel defects. Conclusions from pixel defects are reported as level of support to the hypothesis that an image has been acquired with a specific camera, and/or the level of support to the hypothesis that the have been acquired by a different camera. The following levels of support can be given: no support, limited support, moderate support, strong support, and very strong support. In cases with similar support to both hypotheses, no conclusion can be drawn due to discrepancies. Header and footer-information is often available in the digital files received. The information in the headers and footers is not visible in the image itself, however by using software (for example a hex viewer) the information can be made available. In JPEG images from cameras this information often provides camera settings and brand and type of the camera itself, and sometimes provides information with which software the image has been edited. It is possible to modify the header and footer information by using software, so for forensic casework the examiner has to be aware of this possibility before drawing conclusions. If the header provides information that the image has been taken with a specific camera, it is possible that someone has altered the contents of this header, and that the picture actually has been taken with a different camera. The results of examining pixel defects can be combined with information from the headers and the footers, for determining the integrity and authenticity of images. Several cases that have been received will be discussed in the presentation, and a review is given of the results reported. Image Processing, Pixel Defects, Standard Operating Procedure C20 Preserving Audio Quality When Converting Digital Audio File Formats Kenneth W. Marr, BSEE, MS*, and David J. Snyder III, BSET*, Federal Bureau of Investigation, Forensic Audio, Video and Image Analysis Unit, Engineering Research Facility, Quantico, VA 22135 The goal of this presentation is to describe techniques to convert digital audio files from one format to another and to review precautions and limitations to observe when converting audio files so that the maximum audio quality possible is maintained after conversion. This presentation will impact the forensic community and/or humanity by demonstrating to the digital evidence examiner the knowledge of the procedures and limitations to consider when converting digital audio files so that the maximum audio quality is maintained. 137 Analysis of digital audio files is becoming more and more common for law enforcement in today’s Internet world. Cell phone technology, Internet communications, and commercial digital telephone service are examples of the numerous occurrences in which proper digital analysis procedures and the limitations of digital audio formats must be recognized by the digital evidence examiner. Digital forensic tools must be used with skill, the characteristics of the digital file format under question must be identified, and the proper steps must be followed in order to convert a digital audio file from one format to another while maintaining the maximum audio quality. Selection of proper digital analysis tools will include a forensically approved digital hex editor program; a high quality computer-based audio analysis application, and a high-quality computer and operating system. The work on this presentation is based on a Windows 2000 operating system. The reasons for converting digital audio files from one format to another could include the needs of the requesting agency, errors in the audio file rendering it unplayable, or a variety of many other reasons. The goal of the forensic audio examiner in this conversion is to maintain as much audio quality as possible after the conversion. The format and characteristics of the digital audio file to be converted must be determined. Then the format and characteristics of the converted file must be determined. This can be accomplished by use of a digital hex editor program to identify file characteristics in the header portion of the file. This information could include the type of file, sampling rate, bit rate, Codec, compression characteristics, size of header information as well as the size of the data portion of the file. Comparison of this header information is essential both before and after file conversion. One of the most important features to consider when converting digital audio files is the effect of compression. Since many digital audio files are already compressed when received, case-by-case analysis is required before using any compression algorithms. Nevertheless, preserving audio quality should remain the most important consideration for the digital evidence examiner. Digital Evidence, Digital Audio, File Conversion C21 Case Study - Imaging the Memory of a Digital Audio Recorder Kenneth W. Marr, BSEE, MS*, David J. Snyder, BSET, and Jeffrey Edwards, MSEE, Federal Bureau of Investigation, Forensic Audio, Video and Image Analysis Unit, Engineering Research Facility, Quantico, VA 22135 The goal of this presentation is to review the procedures and precautions for making an exact digital copy of the memory contents of a handheld digital audio recorder. This presentation will impact the forensic community and/or humanity by providing to the digital evidence examiner procedures and precautions for making an exact digital copy of the memory contents of a digital audio recorder. Commercial digital audio recorders are used throughout society today. This case involved a commercial off-the-shelf digital recorder that the submitting agency requested to be examined to determine the contents of the recorder’s memory. Since there were questions involving alleged alterations of the recorded information, an image of the recorder’s memory was required. This is the same concept in use by computer analysis response teams when downloading the contents of a computer’s hard drive. The image of the hard drive is an exact digital copy of the contents of the original drive. This allows the examiner the opportunity to review and analyze the image contents without affecting any parameters of the contents of the original drive. Potentially vulnerable information on a hard drive includes file names and the dates/times files were stored. Since this case involved an unknown recorder system, a thorough search of law enforcement and Internet forensic sources was made to find existing forensic tools which could be used to image the recorder memory. 138 None were found but valuable forensic consultation information was obtained. It was determined that a new procedure must be validated and approved before the image could be obtained. The same make and model recorder with the same characteristics and features was used to conduct the validation testing. Established forensic principles of write-protection, data verification by use of MD5 Hash calculations, and use of validated procedures were followed. Consultation with the recorder manufacturer confirmed the design and operating specifications. Digital tools used in the validation included a forensically approved digital hex editor program, a commercial monitoring utility for USB interfaces, a computer utility written to query the recorder memory contents, and a high-quality computer and operating system. All steps of the procedure were conducted with repeated tests using the same make and model recorder to verify and validate the procedure. MD5 Hash calculations confirmed that the image of the test recorder memory matched the contents of the test recorder. After validation and approval of the procedure, the memory content of the original digital recorder was downloaded and the exact image of the recorder’s memory was returned to the requesting agency. Digital evidence is taking a more and more important role in law enforcement in today’s society. Analysis of digital files requires a high degree of knowledge and expertise to maintain the integrity of the memory contents of digital storage systems. Digital Evidence, Computer Forensics, Computer Image C22 Temperature Tests on Halogen Lamps Harold E. Franck, BSEE, PE*, and Darren H Franck, BSCE, PE, Advanced Engineering, 4713 MacCorkle Ave SE, Charleston, WV 25304 After attending this presentation, attendees will gain knowledge about the dangers and temperatures of halogen lamps and their propensity to start fires. This presentation will impact the forensic community and/or humanity by revealing the fire propensity available when halogen bulbs are used in lamps. Halogen lamps come in a variety of shapes, sizes and wattage options. These lamps are available for a variety of applications including medical and dental, car lamps, as well as household and shop lights. These lamps are filled with halogen gas, which reacts with the lamp filament producing a brighter light source than an incandescent light bulb. Generally, halogen lights are more efficient and have a longer life span than incandescent light sources and are designed to fit most applications. Typical wattage of halogen lamps ranges from 10 watts at 6 volts to 5000 watts at 420 volts. Many of the household type halogen lamps are classified according to their tubular double-ended shape as T-3. This investigation centers on test measurements conducted on T-3 style halogen lamps and a comparison to standard incandescent lamps. A standard incandescent light bulb in the 60 to 75 watt range produces temperatures that vary over the surface of the bulb. At the base the temperature is approximately 120° F while at the maximum radius the temperature climbs to 180° F. This temperature at the maximum radius of the bulb explains why, although very hot to the human touch, a lit incandescent light bulb can generally be unscrewed from its socket and changed. Surprisingly, the temperature at the very top or tip of a standard incandescent light bulb is approximately 350°F. Tungsten-halogen lamps are also incandescent (filament) lamps but are significantly different from standard or conventional lamps in size and design. Halogen lamps operate on the practical application of the halogen regenerative cycle in filament lamps. Iodine and bromine gas, members of the halogen family of gases, are the most commonly used fillers for these lamps. As the lamp burns, the halogen gas combines with the tungsten that is evaporated from the filament. The circulation of the gas inside the bulb deposits the tungsten back on the filament instead of inside the bulb wall. This regenerative effect * Presenting Author keeps the bulb wall clean and allows the lamp to essentially deliver a constant light output throughout its life. In comparison to standard filament lights, halogen lamps may achieve bulb temperatures in excess of 900° F. Consequently, most halogen light fixtures have supposed safety features that isolate the bulb from the surroundings by the use of lenses or guards. In some of these halogen lamp designs, the lenses or guards are insufficient so that significant temperatures may be achieved that will ignite combustible materials such as clothes and paper. Owing to the lamp temperatures, halogen lamps use quartz rather than glass for the lamp bulb. Tests conducted on halogen lamps varying from 150 watts to 500 watts revealed a serious risk of fire on two common light fixture designs. 3.0 4.0 Halogen Lamp, Temperature, Ignition C23 Outdoor Bus Duct - Maintenance, Diagnostics, and Failures Helmut G Brosz, BASc, PEng*, and Peter J.E. Brosz, BEng, PEng, Brosz and Associates, 64 Bullock Drive, Markham, Ontario L3P 3P2, Canada The goal of this presentation is to emphasize the importance of testing and inspection of surviving outdoor bus duct for the forensic engineer; highlight some common manufacturing defects; and show common failure modes and resulting injuries. This presentation will impact the forensic community and/or humanity by demonstrating to forensic electrical engineers faced with outdoor bus duct failures awareness of some newly discovered causes of bus duct failure and the manner of detecting incipient failures. Modern bus duct operating at 120/208/480 and 600 Volts at current rating of 400 – 4000 Amps is generally of the low impedance type, feeder or plug-in-type utilizing one or two bolts per joint type of construction. The conductor is flat ¼’’ thick plated copper or plated aluminum 3, 4 or 5 wire insulated with PVC or epoxy and sometimes with an additional Mylar sheet or tape. The enclosure is often powder coated, painted aluminum or painted steel. The newer versions of bus duct are constructed in a ‘’sandwich configuration’’ and consist primarily of bus bars coated or dipped in either an epoxy or PVC resin. This type of construction has certain benefits and disadvantages. For instance, one draw back is that certain material defects are not readily detectable using commissioning / maintenance test procedures. The bus duct is constructed to one or more of the following Standards UL 857 or CSA C22.2, No. 27. Outdoor bus duct does not accept plug-in switches. Outdoor bus duct is usually installed in a vertical or horizontal manner between transformer and buildings. Outdoor bus duct is expected to be weather sealed except for certain drainage features. 1.0 Failure Modes and Reasons 2.1 A common failure mode is the hot spot fault at a joint or connection. It is often attributed to misalignment, defective manufacture and loose or corroded connections. 2.2 Insulation failure near or at the joints due to defective manufacture, lack of clearances or damaged insulation. 2.3 Damage caused by entry of water, cement, slurry rain and defective weatherproofing. 2.4 Multiple pinholes in the insulation in the presence of water in horizontal installations. 2.5 Bolt hole penetration not waterproofed. 2.6 Incorrect orientation of the bus duct. 2.7 Condensation of moisture. 2.0 Older Bus Duct Older bus duct is often of the higher impedance type with larger spacing between phases. It may be enclosed or ventilated. The * Presenting Author 5.0 6.0 conductors are often supported on separate insulating spacers exposed to rain, snow, etc. Failures have occurred at support points. Injuries Injuries seldom occur on outdoor bus unless being worked on while energized. In these instances, workers are very close to the fault, which usually manifests itself as a sustained electrical arc, which produces intense searing heat along with molten metal ejecta. The incidence of shock and electrocution occurs less often on outdoor bus duct than indoor bus duct. Consequential Damages The most serious outdoor bus duct failures usually result in major building fires and lengthy business interruptions. Sometimes a particular defect exists in an incipient form throughout the entire bus duct system, necessitating shutdown with resulting business interruption in order to take corrective actions on the entire run. Malfunction of protective equipment such as circuit breakers and relays usually increase the degree of damage. Tests Infrared scans, heat runs, insulation resistance, capacitance and dissipation factor/power factor, conductor resistance, plating and insulation thickness, alignment, correct installation, weather and water resistance tests and ‘’holiday’’ tests are a menu of tests often used by the forensic engineer. Case Studies of Bus Duct Failures 6.1 Cement and water slurries 6.2 Tools 6.3 Insulation failure and holidays 6.4 Manufacturing defects 6.5 Hot spots 6.6 Misalignment 6.7 Condensation Electrical Bus Duct, Arcing, Product Liability C24 New Standards in Forensic Engineering & Science Harold E Franck, MSEE, PE*, Advanced Engineering, 4713 MacCorkle Avenue, SE, Charleston, WV 25304; and Helmut G Brosz, BASc, PEng, Brosz and Associates, 64 Bullock Drive, Markham, Ontario L3P 3P2, Canada After attending this presentation, attendees will gain knowledge on the current status of standards available to forensic engineers and the need for further standard development to deal with Frye standard and Daubert challenges. This presentation will impact the forensic community and/or humanity by demonstrating the use of recognized standards in the practice of forensic engineering, which allows the practitioner’s opinions and results to withstand court challenges. Goals of a forensic engineer/scientist include safeguarding the life, health, property, and welfare of the public, establishing and maintaining a high standard of integrity, skill, and practice in the profession of forensic engineering. The rules of professional conduct for engineers dictate that the forensic engineer shall be completely objective and truthful in all professional reports, statements and testimony. He shall include all relevant and pertinent information in such reports, statements, and testimony. When serving as an expert or technical witness before any court, commission, or any other tribunal, he shall express an opinion only when it is founded upon adequate knowledge of the facts in issue, upon a background of technical competence in the subject matter, and upon honest conviction of the accuracy and propriety of his testimony. These statements or similar ones are generally found in the rules of conduct for engineers in the United States and Canada. 139 In light of these requirements of conduct, the forensic engineer can ensure that his opinions, reports, and testimony stand on solid ground if they are based on recognized standards. In the last few years, forensic engineers and scientists have been subject to court challenges as to their opinions. These challenges originally stem from a court case in 1923 that established the minimum standard required for the admission of expert testimony in federal cases. The Frye Standard, as it has become known, requires the expert to use data and methodology “generally accepted” by other experts. A more recent landmark case is the Daubert decision concerning birth defects that were allegedly caused by the mother’s use of the anti-nausea drug, Bendectin. The defendant’s, Merrill-Dow, motion for summary judgment was granted by the trial court because Daubert’s experts relied on a technique that the court felt had not received general acceptance within the scientific community. Furthermore, in support of its finding of no general acceptance, the court observed that the proffered findings had not been published or subjected to peer review. The appellate court affirmed the lower court’s decision based on the Frye Standard. However, the Supreme Court of the United States found that the Federal Rule of Evidence 702, issued in 1975, had superseded the Frye Standard and that “general acceptance” was not the sine qua non of admissibility. Rather, the Supreme Court stated, any reliable and relevant scientific evidence was admissible. For guidance, the Court then listed some suggested reliability criteria, as follows: 1. Has the theory or technique underlying the proffered evidence been tested and found to have a reasonably low error rate? 2. Has the theory or technique been subject to peer review and publication? 3. Are there recognized standards for applying the theory or technique? 4. Does the theory or technique enjoy general acceptance in the scientific community tantamount to the Frye Standard? The Court emphasized that no one of these criteria needed to be satisfied and that they may not apply to all types of scientific evidence. In any event, all federal courts are now governed by this Supreme Court decision. Many states have also accepted the “Daubert Standard,” though there are still a sizeable number that have continued to apply the Frye Standard and a few that follow neither standard. Standards in forensic engineering have two main goals: to educate the practitioner and to codify the recognized practice in a particular field. Consensus standards, such as those promulgated by ASTM, ensure that the procedures followed by the investigating engineer follow recognized and well-documented outlines. These procedures aid the investigator by ensuring that pertinent items are addressed, that available evidence has been gathered, and that a scientifically provable hypothesis has been developed and tested. When properly applied, standards and guides are an invaluable aid to the practicing forensic engineer. These guides also place the investigator’s opinions on solid foundations with respect to the laws of the land as dictated by the Code of Federal Regulations. Various standards, such as American National Standards Institute (ANSI) Institute of Electrical and Electronic Engineers (IEEE), American Society of Civil Engineers (ASCE), Society of Automotive Engineers (SAE), and many others serve to support the forensic engineer’s work and can provide a sound basis for his forensic opinions as well as for having those opinions accepted into evidence at trial. Forensics, Standard Development, Frye Standard C25 Analysis of Case Factors in a Surface Mine Electrocution James A. Ruggieri, PE*, 10710 Timberidge Road, Fairfax Station, VA 22039 After attending this presentation, attendees will understand the application of engineering judgment. 140 This presentation will impact the forensic community and/or humanity by identifying innovative techniques, critical thinking, and application of engineering to judgments and conclusions in forensic sciences. In late November 1999, a 44-year-old dredge operator working a gravel pond from a small metal boat, was electrocuted when he tightened a chain clamp around one phase of an energized 440 Volt, 3-phase dredge service power conductor fed from a corner-grounded delta transformer. The operator was in the process of clamping the dredge rubber discharge line to a metal section of pipe when the power conductor was drawn up with the chain clamp. The operator had a total of two years and four weeks mining experience, all at this mine, as a dredge operator, yet had not received electrical safety and hazard recognition training in accordance with Mine Safety Health Administration regulations (MSHA) 30 CFR Part 48. After struggling for some time to get the chain clamp to seat, the operator hit the chain clamp handle with a pipe to get the chain’s clamping teeth to seat into the chain links. The chain clamp finally seated and the operator began to tighten the clamp. An assistant on the boat reported that he then heard the operator make grunting noises while shaking violently. Recognizing that the operator had contacted a power line, the assistant failed in his attempt to push the operator free, and fearing for his life, entered the water and swam to a nearby floating stanchion. The assistant climbed onto the float and reportedly experienced “tingling” sensation, and when he looked back, observed arcing in several locations. This paper provides an analysis of case factors used in a professional negligence and wrongful death case on behalf of the Plaintiff against two defendants, the mine owner (employer) and an electric contractor tasked in performing annual safety inspections of the mine facility as required by MSHA. This analysis shows that both the employer and the contractor had ample knowledge of substantive electric risk, and provides an axiomatic breakdown of case factors shown to be recurring in such incidents. This analysis evaluates system grounding deficiencies, deficient personnel training and personnel protective equipment policies and procedures, and in general, a grossly deficient system architecture. Additionally, this paper identifies impacts of federal regulatory policies, while evaluating rulemaking politics and enforcement of the provisions of the 1977 Mine Safety and Health Act. Electrocution, Grounding, Surface Mine C26 Forensic Investigation of a Gas Phase Explosion in Building Poh Ling Chia, BSc*, Ming Kiong Michael Tay, PhD, MBA, and Kim Lian Janice Kuah, MSc, Health Sciences Authority, 11 Outram Road, Singapore, 169078, Singapore Through the systematic and detailed examination of the scene and physical evidence collected, the goal of this presentation is to provide the examiner means to identify the fuel source from a number of potential sources and reconstruct the sequence of events that led to the gas phase explosion in a building This presentation will impact the forensic community and/or humanity by illustrating how forensic scientist can complement structural design and civil engineers as well as gas safety experts in determining the cause of gas-phase explosion. Background: The explosion occurred in a design and advertising office on the second floor of a seven-story flatted factory building of reinforced concrete structure with flat roofing. The occupier of the affected unit carried out the manufacture of advertising signs and displays within the premises, which was partitioned into an office area and a production area. Four persons were killed and another two workers were injured in the accident. * Presenting Author Prior to the accident, two of the deceased were involved in the replacement of the oxygen cylinder and the usage of the oxygen-acetylene equipment in the production area. Eyewitnesses reported seeing a dazzling brightness immediately after the initial explosion, suggesting an oxygen-rich fire. Flames were also seen shooting out from one of the liquefied petroleum gas (LPG) cylinders in the office during fire fighting. Scene Examination: The explosion/fire totally destroyed the factory unit. The concrete wall, window frames, doors, and gates at its main entrance were displaced from their original positions. The concrete wall was found to bulge outward, attesting to an explosion from within the factory unit. Along the corridor of the affected factory unit, partially burnt remains of several metal racks, an air compressor (used for spray painting), and a number of paint containers were found. The damages observed to the areas within close proximity to the affected unit included displaced doors of the passenger/cargo lifts, shattered window panes in neighboring units, and broken windows of cars parked downstairs. Within the affected factory unit, partially burnt remains of paint containers and solvents were found at several locations at its production area. Tools such as an electric saw and drilling machine were also found at the scene. At the locality within the production area where the oxygenacetylene equipment was claimed to be used at the time of accident, a circular indentation of about 20 cm diameter was observed on the floor. Also, impact marks were noted on the wall surface nearby, probably caused by fragments from the explosion. A badly dented acetylene cylinder was found, among the debris, at close proximity to the circular indentation. The shattered remains of a second cylinder (believed to contain oxygen prior to the accident) were found at several locations within the factory unit. The damaged remains of two sets of regulators and flexible gas hoses were also recovered from the scene. Two cylinders containing liquefied petroleum gas (LPG) were found within the affected factory unit. No flashback arrestors, non-return valves and torch were recovered from the scene. Potential sources of gas that led to the explosion include: • A leak of flammable gases from the LPG cylinders. • A leak of acetylene from the acetylene cylinder. • The oxygen cylinder and/or hose being contaminated with flammable gases. Findings: (a) LPG cylinders The on/off gas valves of both cylinders were found to be in the close position. The plug within one of the valve was found to be burnt. One of its ends had expanded causing the seat disk within the plug to drop by approximately 0.3cm from the top. This resulted in poor sealing and gas to be leaking out from the on/off gas valve even though it was close. The above findings indicate that both LPG cylinders were not in use before the accident and any gas leakage was the result of heat from the fire. The leaking gas resulted in flames, which were seen to be emitting out from one of the cylinders by the firefighters. (b) Acetylene cylinder, pressure regulator, and red hose The on/off gas valve was found to be in the close position. It was clotted with melted rubbery material from the safety plug within the cylinder. This was consistent with effects from external heating. The pressure release valve was also found to be damaged by the heat from the fire allowing acetylene to escape into the atmosphere. The pressure regulator believed to be attached to the acetylene cylinder was found to be relatively intact. Both the cylinder and working pressure gauges remained fixed to the regulator body. One of the gauges was found to be bent forward indicating external force acting on it. The lengths of red acetylene hoses had damaged ends and were found to be intact with burn marks along their lengths. These damages were consistent with those caused by external forces and heat. The above findings indicate that the acetylene cylinder was not in use before the incident and any gas leakage from the cylinder was the result of heat from the fire. * Presenting Author (c) Oxygen cylinder metal fragments, pressure regulator, and blue hose The on/off gas valve was found to be in the open position. Four of the metal fragments physically fitted to form the base of the cylinder with a diameter of approximately 20 cm. This was consistent with the size of the indentation mark found at the scene. One of the fragments was found to bend outwards indicating an outward force from within the cylinder. The pressure regulator believed to be attached to the oxygen cylinder was found to have its regulator bonnet and working pressure gauge blown off from the regulator body. Soot and burn marks were observed within the bonnet and regulator indicating internal burning within the regulator. The blue oxygen hoses were severely damaged with splits and burn marks along its lengths consistent with those caused by internal bursting forces and heat. Damages observed on the damaged oxygen cylinder metal fragments, pressure regulator and blue hose indicated that the cylinder was used during the time of incident and that flashback had occurred along the lengths of blue hoses causing them to split. Burning and excessive pressure within the regulator caused the bonnet to be detached and the working pressure gauge to be blown off. A powerful outward force from within the cylinder caused it to fragmentize. The above findings indicate that both the LPG and acetylene cylinders were not in use prior to the explosion. The source of gas that led to the explosion was hence likely to be from the oxygen cylinder and/or hose. Possible causes for the explosion: 1. Contamination of the oxygen hose by flammable gases from paint cans, acetylene or other sources to within explosive limit resulted in an explosion when the torch was ignited. 2. Contamination of the oxygen cylinder by flammable gases. 3. Absence of safety devices e.g. flashback arrestor, non-return valves in the oxygen-acetylene gas equipment resulted in the explosion of the oxygen cylinder. Typical set-up and operation of oxygen-acetylene equipment: The oxygen-acetylene equipment was primarily used for cutting, welding, brazing, or heating of metals. A typical set-up of oxygen-acetylene equipment comprises oxygen and acetylene cylinders, gas regulators, flashback arrestors, hoses, non-return valves, and a torch. A gas regulator serves to regulate/reduce the pressure of the gas coming out from the cylinder before feeding to the hose/torch. A flashback arrestor prevents the flashback of in the torch/hose from propagating back into the regulator and gas cylinder, which could lead to an explosion. The function of a nonreturn valve is to prevent the gas from flowing back into the cylinder. The internal pressure of an oxygen cylinder (when full) is around 150 bar and the internal pressure of an acetylene cylinder (when full) is around 20 bar. The two types of gases are channeled via hoses to a torch where the gases are mixed and ignited. The low-pressure control valves are typically adjusted to 2 bars for oxygen and 0.2 bars for acetylene before the valves are opened to release the gases into the hoses. The oxygen valve at the torch was then opened momentarily to clean the nozzle and closed back. The acetylene valve (about ½ turn) will then be opened, followed by the oxygen valve (about ¼ turn). The torch is then ignited. To stop the torch, the torch’s acetylene valve was closed followed by the oxygen valve in order to prevent acetylene from flowing into the oxygen hose. Due to the potential fire hazards associated with oxygen-acetylene equipment and the strict protocol required in its operation, only trained personnel should be allowed to operate the equipment. Tracing the source for contamination: Experiments conducted on an oxygen-LPG equipment not equipped with non-return valves or flashback arrestors showed that if the internal pressure of the LPG cylinder (when full) was higher than that of the oxygen cylinder (when it was near empty), and if the torch nozzle was obstructed, it was possible for the LPG to flow into the oxygen cylinder. The oxygen in the oxygen cylinder would thus be contaminated with LPG. Investigations showed that no non-return valve and flashback arrestor were connected to the oxygen-acetylene equipment used in the affected 141 company. Also, the company was not licensed to carry out hot works within the office premises; hence the workers might not be trained in the operation of oxygen-acetylene equipment. The absence of these safety devices coupled with untrained workers using the equipment may have resulted in acetylene contaminating the oxygen hose and cylinder resulting in the explosion. Gas Phase Explosion, Oxy-Acetylene Gas Equipment, Flashback Arrestor C27 Lethality of Taser Weapons James A. Ruggieri, PE*, 10710 Timberidge Road, Fairfax Station, VA 22039 Upon completion of this presentation, attendees will understand applications of some engineering applications. This presentation will impact the forensic community and/or humanity by identifying innovative technologies, critical thinking and application of engineering to judgment and conclusions in forensic science. On April 28, 2004, Montgomery County Police MD police responded to a disturbance. A 45-year-old mentally ill adult, 6’4”, 275-pound son pushed his elderly mother when he became agitated when a car carrying Chinese delivery food parked in front of the suspects house. The driver left the headlights on and ran next door to deliver food. Family members said the suspect apparently believed the car carried agents comi
